ABSTRACT

HAIRBASE: THE DEVELOPMENT OF AN ONLINE REFERENCE ATLAS OF MAMMALIAN HAIR

The class Mammalia contains approximately 5,400 species that display incredible variation in their pelage characteristics. When attempting to identify an unknown animal based on the morphological characteristics of its hair, the macroscopic and microscopic examination of guard hairs is often used. Species identification from small amounts of hair or single hairs has been the goal of many published atlases. However, many of these atlases do not adequately reflect the range of variation both between and within individual species. HAIRbase™ is an online digital database of bright field and scanning electron microscope images of mammalian hair. In contrast to many other atlases, HAIRbase™ contains images from both primary and secondary guard hairs from three distinct regions on the body from each species represented. HAIRbase™ is an applicative tool that can aid forensic hair examiners by providing a visual reference that contains diagnostic information regarding traits of the hair shield, subshield, and basal regions, including medullary configurations, cortical characteristics, and cuticular patterns that can be used to generate leads in investigations involving animals.

Michael Vincent Gonzalez May 2011

HAIRBASE: THE DEVELOPMENT OF AN ONLINE REFERENCE ATLAS OF MAMMALIAN HAIR

by Michael Vincent Gonzalez

A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Forensic Science in the College of Science and Mathematics California State University, Fresno May 2011

© 2011 Michael Vincent Gonzalez APPROVED For the Department of Chemistry:

We, the undersigned, certify that the thesis of the following student meets the required standards of scholarship, format, and style of the university and the student's graduate degree program for the awarding of the master's degree.

Michael Vincent Gonzalez Thesis Author

Kevin W.P. Miller (Chair) Chemistry

James P. Prince Biology

Bonnie Yates National Fish and Wildlife Forensic Laboratory

Margaret E. Sims National Fish and Wildlife Forensic Laboratory

For the University Graduate Committee:

Dean, Division of Graduate Studies AUTHORIZATION FOR REPRODUCTION OF MASTER’S THESIS

X I grant permission for the reproduction of this thesis in part or in its entirety without further authorization from me, on the condition that the person or agency requesting reproduction absorbs the cost and provides proper acknowledgment of authorship.

Permission to reproduce this thesis in part or in its entirety must be obtained from me.

Signature of thesis author: ACKNOWLEDGMENTS This thesis has been a long and arduous journey beset with many difficulties. As it stands complete, I can only say, thank you to my strong support system of family, friends, and co-workers that not only helped make this project the success that it was, but also helped me keep my sanity during those tough days in the laboratory. First, I would like to thank my mother and father, without them, I would not be here (both literally and figuratively). I would like to thank my father for instilling in me the benefit and importance of a strong education. Also, I would like to thank him for always being in support of any crazy dream that seemed to pop into my head (even if it meant moving to Fresno to pursue my Master’s degree in Forensic Science). I would also like to thank my mother, who has been a pillar of support through good times and bad. She has always been the one reminding me to stay humble, even in the face of educational success. I would like to thank her for genuinely believing that I can do anything that I set my mind to, and then convincing me to go and do it! I would also like to thank my twin sister Casey, a large part of my cheering section, and someone who thinks the world of me. Always calling me the “smart twin” (despite her own educational success) and always being a willing ear to listen in any circumstance. She has been my voice of reason through all the trials and tribulations of graduate school (and life for that matter). Honest to a fault, she has always been ready with what I needed (not always what I wanted) to hear. I don’t believe that I could have made it through without her in my corner. I would like to thank and express my deepest gratitude to Ms. Elsbeth Murata – my right hand in the laboratory and also one of my best and closest vi friends. It would be an understatement to say that she merely “helped” me with this project. She took equal part in the development of HAIRbase™, often willing to give more of her time and resources to this project than I was. Without her, this project would not be half of what it is today. I have seen her grow from an inexperienced undergraduate into quite an accomplished hair morphologist in her own right. I know that she will go on to do great things in the sciences. She has been one of my biggest supporters and always knew that I could do it (even when I doubted myself). I want to thank her for not only putting so much tireless effort into this project, but also for putting up with me (way more than she probably wanted to). Hours of imaging of individual hairs, marathon photo-shopping sessions that went into the wee hours of the morning, she was there through it all (and to my surprise, didn’t want to leave!). I know that a mere thanks in an acknowledgement section of a thesis does not encompass the amount of credit that she deserves, but again, thank you so much! I would like to personally thank Maria Carrizales for her tireless efforts to buoy my spirits after long, rough days and being a strong support system that I could lean on in times of trouble. She was always ready at a moment’s notice to edit any writing, listen to a speech, or answer any question that I had (despite her own work dealing with her PhD). She is the definition of a true friend. I would like to thank her for always being there, and also, for giving me “tough love” when I really needed it. I would like to thank my thesis committee for their efforts to keep me on track and staying positive. I would like to thank Bonnie Yates and Cookie Sims for teaching me not only the finer points of hair morphology and trace analysis, but also how my project would function in the professional realm. They always believed that I could finish and that I would go on to do great things! Both of them vii were always in my corner and were a large part of my cheering section, even when deadlines were looming and things looked their bleakest. I would like the thank Dr. Prince for being my “non forensic committee member”. Approaching the project from a strictly biological sense gave the project more depth and made it a more valuable resource. I want to thank him for his efforts as an editor and as a great source for feedback of a different perspective. I would like to thank Dr. Kevin Miller for standing by my side during every iteration and troubleshooting endeavor that came with this project. I want to thank him for always making himself available should the need arise and planting it in my head that I had what it took to get my PhD. He has helped me, not only to succeed professionally, but also in my personal life as well. He has been there through thick and thin (kidney failure being a particularly “thin” time). He has been one of my biggest cheerleaders throughout my graduate school journey. I also wanted to thank him for pushing me to be more than even I thought I could be, he has truly been instrumental in my success. I would like to thank the California State University, Fresno Biology Department and the National Fish and Wildlife Forensic Laboratory for donating hair samples to this project. To all the other people (you know who you are) that were there to help me out and encourage me, I give a very sincere thanks!

TABLE OF CONTENTS Page

LIST OF FIGURES ...... x

INTRODUCTION ...... 1

LITERATURE REVIEW ...... 6

Hair Physiology ...... 6

Macroscopic Variation of Hair ...... 8

Microscopic Variation of Hair ...... 10

Hair Atlases and Classification Schemes ...... 12

MATERIALS AND METHODS ...... 17

Sample Collection ...... 17

Loose Hair Storage ...... 18

Preparation of Hair for Microscopical Examination ...... 19

Microscopy Slide and Stub Storage ...... 21

Digital Image Acquisition ...... 21

Photo Manipulation ...... 24

Development of an Internet-based User Interface ...... 25

RESULTS AND DISCUSSION ...... 26

Development of HAIRbase ...... 26

User Guidelines ...... 34

Future Research ...... 35

SUMMARY ...... 38

REFERENCES ...... 39

APPENDICES ...... 44

APPENDIX A: SPECIES INCLUDED IN HAIRBASE™ ...... 45 ix Page

APPENDIX B: WEBPAGE TEMPLATES ...... 52 APPENDIX C: HAIRBASE™ IDENTIFICATION NUMBERS AND CONGENERS ...... 57 APPENDIX D: HAIRBASE™ SPECIES COLLECTION AND STORAGE CHECKLIST ...... 65

APPENDIX E: HAIRBASE™ MICROSCOPE PREPARATION CHECKLIST 74 APPENDIX F: HAIRBASE™ IMAGING AND WEBPAGE DESIGN CHECKLIST ...... 81 LIST OF FIGURES

Page

FIG 1: The degree of variation between the grades of hair on Lycalopex griseus (Argentinian Grey Fox)...... 3

FIG 2: The structure of primary and secondary mammalian guard hairs...... 5 FIG 3: Petraco and Kubic’s (7) identification scheme for unknown mammalian hair...... 13 FIG 4: Brunner and Coleman’s (10) identification scheme for unknown mammalian hair...... 15

FIG 5: Teerink’s (3) identification scheme for unknown mammalian hair...... 16 FIG 6: Hair regions collected: Hair was plucked with tweezers from the dorsal region (3), ventral region (4) and tip of tail region (5) of each specimen...... 18 FIG 7: Loose hair storage and organization: A. represents an overall image of the three-ring binder dividing the collected species. B. Picture of the large Ziploc bag with labels. C. Small plastic bag for individual region on each specimen...... 19 FIG 8: Bright field microscopy slide storage. Microscope slides with whole mount specimens are on the left with a comprehensive listing of the slides on the right...... 22

FIG 9: HAIRbase™ image template...... 24

FIG 10: Cellular classifications of the mammalian medulla...... 29

FIG 11: Cuticular pattern classifications of mammalian hair...... 30

INTRODUCTION

Illegal hunting and the fur trade comprise a $4 billion dollar black market industry, second in the world only to the trafficking of narcotics and weapons (1). The identification of trafficked wildlife may rely on accurate diagnosis of hair left behind at a crime scene, as trace evidence found on objects connected to illegal activity, or from body parts that are themselves the trafficked items. Hair may come from a variety of mammalian species (2-4), each of which demonstrates a great deal of morphological variation that can aid in the identification of the animal from which it came (5-7). Therefore, the analysis of mammalian hair morphology is an important tool that wildlife forensic scientists and animal welfare professionals can use to combat these illegal activities (8-10). Hair morphology has been studied in academic circles for many years (3,5,7,10-14). Hair morphological studies are vital to forensic professionals attempting to make taxonomic designations based upon the identification of hair fiber characteristics entered as evidence or found at a crime scene (3,5,7,10). Although reference materials play an important role in any forensic case that relies on morphological comparison, it is important to remember that comparative studies ultimately rely on the use of a verified hair reference standard collection. One should always compare like to like: that is, to ensure that standard and questioned hairs are comparable in their macroscopic features (e.g., length, diameter, color, banding pattern) and type (e.g., guard hair, secondary, underhair) in the first instance. Then, it is important to compare similar portions of one hair to another (e.g., basal to basal, shield to shield). Animal hair atlases have proven invaluable to forensic professionals by illustrating morphological differences that 2 can provide a fast and efficient way to eliminate certain species while suggesting others that may require further investigation (3,5,7,10-14). Over the years, several atlases of hair morphology have been published that provide representative images of the degree of variation among hairs in order to aid in the identification of certain mammalian groups (3,5,7,10-14). These atlases have been of great use to comparative hair morphologists over time. However, current atlases are either limited in their geographical scope (e.g., Europe, Wyoming, Australia) or lacking in the degree of morphological variation that can occur both within and between animals when the animal and/or body part of origin or the country of origin of the evidentiary hair is unknown. Many laws, including the Endangered Species Act of 1973 (15) and the Marine Mammal Protection Act (16), and trade agreements, such as the Convention on International Trade in Endangered Species of Wild Flora and Fauna (CITES) (17,18) determine which mammals are protected and to what extent these animals are protected. Many published mammalian hair atlases do not take endangered and trafficked animals into account and, so, their utility to the forensic wildlife hair examiner is often limited. Likewise, few of these atlases offer full coverage of the range of variation that may be seen among domestic animals, and which must be taken into consideration during a forensic examination. Current atlases are further limited by the amount of variation they record across the different body regions of a given individual of a specific species. The class Mammalia is extremely diverse, comprising roughly 5,400 species organized into 29 orders (19). As a result of the level of morphological diversity encountered across all mammalian hairs, guard hairs from the dorsal region of an individual (medially between the shoulder blades) have been the hairs most commonly 3 referenced (3,5-7,12,13,20). However, a large number of mammals, including furbearers and other endangered species, show variance in pelage characteristics from one body region to another (21,22)(Fig. 1).

FIG 1: The degree of variation between the grades of hair on Lycalopex griseus (Argentinian Grey Fox). Hairs from the back, or dorsum (A), differ from hairs of the tail, or caudal region (B) in length, color pattern, and degree of coarseness. Both of these types of hairs, in turn, differ markedly from the hairs of the belly, or ventrum (C).

A forensic animal hair atlas must also address the variation in diameter or “grades” of hair that may be observed on a particular body region of a given individual. A mammal’s coat contains several grades of hair which are separated into two broad categories, underhair and overhair (3,23,24). Underhair mainly functions to insulate the mammal and provide warmth (3,23,24). Underhairs have been found to be morphologically simple, offering few characteristics that can be 4

used to distinguish species and, so, are of limited value to the forensic examiner (3,10). Overhair, on the other hand, includes the primary and secondary guard hairs, which function to provide protection (5, 9,11,15). True overhairs, sensu scricto, are those extra long hairs, found primarily in furbearers scattered sparsely over the body and are generally too large in diameter and undifferentiated to be useful in microscopic identification. These true overhairs are termed GH0 in Teerink’s scheme of grading mammalian hairs (Fig. 2A (3)). The overhairs that contain the greatest diagnostic value include the primary guard hairs (GH1) and secondary guard hairs (GH2)(Fig. 2A (3)). Primary and secondary guard hairs are morphologically distinct and are, therefore, of particular interest to forensic hair examiners (3,6,10). The cuticle, cortex, and medulla of primary guard hairs, especially from the dorsum, have distinct characteristics that can be used for identification purposes (6,7,13,14,22,25,26). Secondary guard hairs form gradations between the longer primary guard hairs and the shorter, more dense, finer underhair or fur (3,10). The morphological characteristics of secondary guard hairs are oftentimes more diagnostic than those of primary guard hairs (27). However, despite this fact, secondary guard hairs are not taken into account by most hair atlases. Lastly, most current atlases are available in textual form only, including CD-ROM versions (12,28), making the representative image collections static by nature. However, the needs of the forensic wildlife hair examiner are closely aligned with the international judicial system (29,30), and change depending on the current standing of endangered species litigation (29,30). HAIRbase™ is a web-based reference atlas, specifically geared toward the needs of the forensic trace professional. This database attempts to alleviate many of the shortcomings of previous hair morphological references. These 5 shortcomings include species selection relevant to the forensic examiner, geographic scope, limits in the coverage of variation on a specimen, and the static nature of these primarily textual references. HAIRbase™ attempts to alleviate many of these issues by including broad inclusion of forensically relevant species, a greater geographic coverage, a wider representation of morphological variation among and within individuals of the class Mammalia, and the adaptability that digital media provides. By addressing these problems with existing atlases, HAIRbase™ can become an important applicative tool to trace examiners in multiple scientific disciplines for years to come.

FIG 2: The structure of primary and secondary mammalian guard hairs. (A) Primary and secondary guard hairs (GH1 and GH2, respectively) are morphologically distinct, so they are of particular interest to forensic hair examiners. (B) The types of primary and secondary guard hairs that were chosen for inclusion in HAIRbase™ (top) and the morphological regions – namely the shield, subshield, and basal – of each hair that for which images were taken for inclusion in the database (bottom) are illustrated. This figure was adapted and redrawn from Moore, et al., 1974 (5) and Teerink, 1991 (3).

LITERATURE REVIEW

The morphology and structure of mammalian hair has been studied for many years (3,5,7,10-14). The information and literature regarding the development, structure and function of hair can fill many volumes and the information, in its entirety, cannot be covered here. However, the use of certain characteristics and structures of mammalian hair for identification purposes is a relatively novel idea (22). The search for identifying characteristics of mammalian hair began in earnest in the early 1920’s and has continued to the present day (9, 31-33). From this beginning, a number of proposed techniques and reference atlases that attempt to increase the reliability of this comparative science have been produced (3,5,6-7,10,34). Although, the techniques applied are different due to technological advances in microscopy for example, many of the basic tenets of comparative hair morphology for identification purposes remains the same. With recent litigation regarding the protection of mammalian species (29- 30), the protection and conservation of these animals has come to the forefront in recent years. Often, in a wildlife forensic context, examiners are not required to make identifications down to an individual animal but instead are expected to make a taxonomic designation at the species level. Due to the variation of the hair morphology in the class Mammalia, this is often difficult. However, the search and classification of these identifying characteristics has proven instrumental in the prosecution of crimes against animals and still remains a valuable tool in the forensic examiner’s arsenal (3,5,7,10,34).

Hair Physiology The hair follicle is truly a remarkable structure, complete with muscles (the arrectores pilorum), a rich blood supply, nerve endings (making hair a valuable 7

vehicle for sensory response and tactile sensation), and a number of glands that perform duties ranging from waterproofing to hair cycle regulation (24,35-37). It is an organ – developed in utero as an invagination of the epidermis – in which division, differentiation, and migration of cells occur (24,36). Different processes in the hair follicle give rise to the growth of the hair fiber (36-37). This activity results from the hardening of the contents of the medullary, cuticular and cortical cells of the hair shaft (24,36). The hair fiber is the major component of the hair follicle. A hair is characterized as a long thin cylinder of keratinized cells, which contains three cellular components (36-37). The life cycle of hair is a dynamic, continuous process composed of three phases: anagen, catagen and telogen (37). The hair cycle is a pattern of growth, regression, and loss of the hair fiber (24,37). This process of hair cycling enables animals to adapt to drastic changes in environmental conditions (i.e., changing seasons). The first phase of the hair cycle, known as the anagen phase, is characterized by a period of high metabolic and mitotic activity, which initiates the growth of the hair fiber (36-37). The next phase of the hair cycle is the catagen phase, during which the follicle undergoes gradual and orderly morphological and functional changes as it enters its regression phase (24,37). The hair then ceases to grow, where it usually stays in a suspended state for about 2 to 3 weeks (36-37). The final stage of the hair cycle is the telogen phase, also known as the mature stable state of hair growth. The telogen phase can last up to 4 months and the hairs found in this phase have a very short follicle (24,37). Hairs found in this phase are most often shed and this phase is only finished when a new anagen phase is commenced (24,37). These telogen hairs are most often found at crime scenes and are commonly used in criminal investigations (24). 8 Macroscopic Variation of Hair The evaluation of a questioned guard hair begins with a macroscopic evaluation. Macroscopic characteristics that are particularly important to note are color, banding pattern, length, and form of the hair fiber in question (3,5,7,10). The form of the hair refers to its overall shape. Hair can vary in both shape and structure; however, there are regions on each hair fiber that are generally recognized. The basal portion of the hair contains the root end, usually comprising about one-fifth of the hair shaft (5,7,10). The subshield is characterized as the area immediately below the shield region (if present) and comprises the middle portion of the hair shaft (5). The shield region is a widened, flattened area located at various positions on the more distal portion of the hair shaft (5). Hair may show extreme variation along the length of the hair shaft, making the representation of each region important to illustrate the variation that can be seen on a given species. Hair diameter determines the various grades of hair and includes, in decreasing diameter: vibrissae, bristle hairs, overhairs, guard hairs, and underhairs (3,10). The ability to recognize the different grades of hair is an invaluable skill, which can only be developed by repeated exposure, such as that commonly experienced by the forensic trace evidence examiner. Vibrissae, which are commonly referred to as whiskers or tactile hairs, are large, stiff hairs that function primarily as sensory organs (10). Vibrissae are thick hairs that do not lend themselves to readily to transmitted light microscopy and, so, are seldom studied. Indeed, like coarse overhairs, vibrissae may not exhibit traits that facilitate their identification at lower taxonomic levels. Also, vibrissae are relatively few in number on a given carcass, and they are not frequently left behind at the scene of a crime. 9

Bristle hairs are rather stiff, rigid hairs that are specialized in some mammals (10), such as the domestic pig. Bristle hairs are generally consistent in diameter along the length of the hair shaft (10). The medullae of these hairs are generally very thin to absent. Bristle hairs commonly exhibit frayed tips. Most mammalian pelages show the presence of coarse overhairs, which are sparsely scattered across the coat of the specimen (3,10). These overhairs are noticeably longer than the guard hairs (10). These hairs, like vibrissae and bristle hairs, have little microscopic value at lower taxonomic levels. As a result, course overhairs were not chosen for inclusion in HAIRbase™. Guard hairs are the next largest hairs in size, and these hairs comprise a large proportion of the pelage of most mammals (3,5,7,10-14). Guard hairs are of traditional importance in hair identification, because they have been seen to display the most diagnostically relevant features (3,5, 7-8, 10-14). It is also important to note that there are a variety of sizes or grades of guard hairs that also require recognition, separation, and separate scientific analysis. Finally, underhairs are shorter and finer than guard hairs, and they commonly have an undulating appearance (3,5,10). Underhairs remain fairly consistent in diameter along the lengths of their shafts. Underhairs can be similar in structure and function among many mammalian taxa, so they may be of only limited diagnostic use to the forensic trace evidence examiner seeking identification below the taxonomic level of family. It is important to note that, on most furbearing animals, a continuous gradation of hair fibers exists from one hair “type” to another which often makes the separation of the different hair types difficult on these animals (10). However, the ability to effectively separate the different grades of hair from one another can aid in the successful identification of a given animal. 10 Microscopic Variation of Hair Many microscopic morphological features of hair have been used to identify the type of animal from which the hair originated (5,10-14). As mentioned above, hair cells differentiate into three layers – the cuticle, medulla, and cortex – and each layer has a distinct function that contributes to the morphology of the hair fiber (24). Identification of the patterns of these differentiated cells often form the basis of forensic hair identification in both humans and non-humans (14,23- 24). The outermost layer of hair, known as the cuticle, is composed of flattened scale cells (scales that overlap) which surround the hair completely, offering protection to the more delicate cortex (3). Cuticle cells grow over one another in an overlapping fashion from the hair’s root to its tip (3,7,38-40). The cells that comprise the hair’s cuticle overlap to form recognizable patterns that are indicative of certain mammalian orders and families (7,38-40). In some rare cases, these scales may be pigmented, as seen in some bats (10). Along most of the hair fiber, scales are flattened against the body of the hair; however, towards the basal region, these scales may be more apparent under bright field microscopy (5). The cortex is composed of spindle-shaped cells that measure approximately 80-100 μM long by 5-10 μM wide (24). The chemical bonds found within the cortex contribute to the overall strength of the hair, while the cortical pigments are responsible for giving hair its color (10,24). Under the light microscope, the individual cells of the cortex are not visible. Rather, they appear as a hyaline mass without much detail. For this reason, the cortex is limited to pigment characteristics that may be useful for identification purposes (3,5,7,10). The medulla appears as a central column of cells that runs medially through the hair shaft (6,9,10,23,32). A medullary structure is not always found within 11 mammalian hair. However, when it is present, it can be classified in a number of ways (3,7,10). The primary purpose for the medulla is presently unknown, but common hypotheses include increasing the thermal insulating properties of hair, forming a channel for waste removal, and maintaining the diameter of the hair (24). The cellular composition and diameter of the medulla varies widely across the class Mammalia (10-11) and, therefore, may be used to help identify a questioned animal hair. Hair morphology remains an important facet of forensic analysis, especially regarding the prosecution of crimes against wildlife (3,5-6). Morphology is simply the study of shape and structure, and this is helpful in a forensic context, because the identification of morphological characteristics can be used to make taxonomic determinations. The identification of trafficked materials often relies on the accurate morphological analysis of hair fibers found at a crime scene, confiscated at customs, or found on objects that are connected with illegal activity. The proper and accurate diagnosis of these hair fibers relies on many morphological features that, together, may make a taxonomic identification possible (5,7,10). Reference materials play a vital role in the analytical process regarding the evaluation of unknown hair samples. The organization, classification scheme, and dichotomous keys devised for reliable identification of unknown animal hairs differs from reference to reference (3,5-7,10). These differences in the ways that morphological structures are described often make it difficult to use these references to their fullest. The biggest benefit of reference atlases to the forensic trace evidence examiner is in the range of morphological variation that the given atlas is able to display. Although no hair atlas or database will ever be complete, shortcomings can certainly be addressed and improved to the benefit of the forensic trace evidence examiner and the profession as a whole. 12 Hair Atlases and Classification Schemes Researchers have long recognized the benefit of hair morphology references evidenced by the number currently available in circulation (3,5,7,10,22,23). These references are primarily textual, and they attempt to offer adequate coverage of the morphological variation seen within the class Mammalia. They are often organized using a geographical scope, cataloguing the species of a particular state or region of a country. This substantially narrows the number of species that need to be addressed and helps to make the construction of dichotomous keys much simpler. However, many times, especially in a forensic context, the origin of a particular (“questioned”) hair fiber is not known. The application of forensic unknowns to geographically-based references limits their utility. In order to illustrate the limited utility of geographically-based references, let’s look at four atlases of mammalian hair identification (3,5,7,10). The identification schemes of each of these texts differ markedly, not only in the methodologies employed, but also in the naming designations of the morphological structures under view. The texts also differ in geographic region covered, number of images displayed, and opinion regarding the diagnostic relevance of certain methods. In fact, the successful identification of an unknown animal hair cannot be applied to any one specific methodology, but must rely on the aggregation of many characteristics including a thorough macroscopic and microscopic evaluation. No one specific method is superior to another. Rather, all characteristics that can be gathered are beneficial in order to make a reliable taxonomic determination based on the morphological characteristics of a given animal hair. 13

Nicholas Petraco and Thomas Kubic (7) offer an identification scheme for unknown mammalian hair. The classification scheme begins with the analysis of cuticle scale casts to view the cuticle scale pattern of the basal portion of the animal hair in question (7) (Fig. 3). Then, classification of the cellular composition of the medulla is completed (7). These two observations are reported by the author to be an adequate means of separation for common animal hair encountered by professional hair examiners. Although this identification process may prove satisfactory for a small subset of mammals, when applied to the entirety of the class Mammalia, it cannot designate lower taxonomic levels. A taxonomic designation cannot be based solely on two analytical methods in order for a reliable identification to be made.

• Imbricate •Mosaic •Petal Cuticle Scale • Diamond Petal/Pectinate Casts •Chevron

• Amorphous • Lattice •Aeriform Medullar • Globular Classification • Multiserial Ladder

FIG 3: Petraco and Kubic’s (7) identification scheme for unknown mammalian hair. This identification scheme requires classification of the cuticular pattern followed by the medullar classification in order to make an identification of an unknown mammalian hair. 14

Brunner and Coleman (10) offer a different approach to the identification of the hair of a number of mammal species. Whereas Petraco and Kubic (7) relied on the analysis of the cuticular pattern as a launching point for further investigation (7), Brunner downplays the importance of such a technique saying, “scale patterns can very often be ignored in the identification process and are generally only used to confirm identifications made on the basis of other criteria”(10). Brunner speculates that the transverse cross-sectional shape of the hair fiber should be the first identifying feature, followed by medullary cellular analysis and designation (10). The naming designations of the cuticle and medulla, while similar, have some differences between the two atlases as well (see Figs. 3 and 4). By viewing these two very different identification schemes, one may begin to appreciate the difficulty of animal hair identification. The difficulty of animal hair identification relies on the premise that the naming and designation of certain morphological structures of hair are subjective, or contingent on the analysis of the individual trace evidence examiner. The identification of individual structures can differ from examiner to examiner, which can often compound the problem of forensic identification of animal hair fibers. The final two atlases have no discernable scheme for identification. Rather, they advocate the importance of an aggregation of characteristics for the proper identification of an unknown hair fiber. Teerink (3) and Moore and co-authors (5) have written atlases that differ markedly in regional scope, but approach hair analysis in much the same way. These two atlases realize the importance of the macroscopic examination of unknown hair fibers and attempt to separate species initially using this criterion. Petraco and Brunner have different opinions regarding the efficacy of certain analytical tests to identify hair. Petraco is a proponent of cuticle scale casts as an initial means of separation of unknown animal hairs, while 15

FIG 4: Brunner and Coleman’s (10) identification scheme for unknown mammalian hair. This identification scheme requires cross-sectional classification, then medullar classification, and finally cuticle scale classification in order to identify an unknown mammalian hair.

Brunner relies on the transverse cross-sectional shape of said animal hairs (7,10). Neither Teerink nor Moore and co-workers comment on the importance of any particular test or structural feature but, instead, they rely upon the aggregation of multiple characteristics including macroscopic observation, cuticular pattern classification, and medullar classification (3,5). In addition, Teerink investigates the cross-sectional shape of the hair fiber (3) (Fig. 5). By analyzing this small subset of reference literature, differences in classification schemes (e.g., medullary types, cuticlular scale types) and naming designations that often – as well as the importance that Brunner and Petraco place on certain analytical techniques for the purposes of hair identification – can begin to be appreciated (3,5,7,10). Therefore, it is a number of characteristics rather than a schema that involves only one or two characteristics that will ultimately lead to the successful identification of an unknown animal hair. No analytical technique is 16 more important than the next, and all techniques should be used in tandem to make a successful taxonomic designation based on the morphological characteristics of an unknown mammalian hair.

FIG 5: Teerink’s (3) identification scheme for unknown mammalian hair. The identification of an unknown mammalian hair requires the aggregation of all characteristics (see Appendix F for Classification types that are possible).

MATERIALS AND METHODS

Sample Collection Animal specimens were obtained from the collections at the U.S. Fish and Wildlife National Forensic Laboratory in Ashland, Oregon, and the Biology Department at the California State University, Fresno, representing 133 species from 19 families representing 2 orders of the class Mammalia. Of these, 46 species are either endangered or protected (Appendix A). Hair was collected from each specimen by either plucking it or cutting it as close to its base as possible with a razor blade. Hair was collected from three body regions: 1) the dorsal region, between the shoulder blades; 2) the ventral region, on the midline between the forelimb and the hind limb; and 3) at the tip of the tail (Fig. 6). For animals that did not have prominent tails (e.g. the chimpanzee, Pan troglodytes), hair was collected from the sacral region of the specimen. These three body areas were chosen, because they provided the greatest level of variation within a given individual while being manageable for a large number of species. Animals that exhibited sudden or marked differences in appearance over their entire bodies (e.g. the Okapi, Okapi johnstoni) were sampled in the additional body regions: back of thigh, forehead, top of foot, and vibrissae. Approximately 20-25 hairs were collected from each body region of each individual, for a total collection of approximately 60-75 hairs from each animal. The collected hair from each body region was then placed in a separate sterile biohazard plastic specimen bag (Fisherbrand, Part # 01-800-00). The plastic specimen bag was sealed and labeled with the following information: genus, species, common name, area on the body from which the hair was collected (i.e. dorsal, ventral, tip of tail, back of thigh, forehead, top of foot, or vibrissae), and sample provenance (if known).

18

FIG 6: Hair regions collected: Hair was plucked with tweezers from the dorsal region (3), ventral region (4) and tip of tail region (5) of each specimen.

Loose Hair Storage The collected hair samples were organized and stored in the Human Identification Laboratory for future use. The plastic specimen bags of collected hair from the selected regions on each specimen were placed in standard sheet protectors with a backer card displaying the common and scientific name. These species “packets” (including plastic specimen bags containing hair from the separately collected body regions and provenance information) (Fig. 7B, 7C) were then organized into individual three-ring binders (Fig. 7A). The three-ring binders were divided by the taxonomic order (e.g., Carnivora, Artiodactyla, etc.). The individual species belonging to that particular order were further arranged alphabetically by family (Mustelidae, Procyonidae, etc.). The representatives within each family were sorted alphabetically by genus. This organizational system was used to facilitate fast and efficient navigation through the loose hair collection. 19 A B

C

FIG 7: Loose hair storage and organization: A. represents an overall image of the three-ring binder dividing the collected species. B. Picture of the large Ziploc bag with labels. C. Small plastic bag for individual region on each specimen.

Preparation of Hair for Microscopical Examination

Preparation of Hair for Bright Field Microscopy Hair from each body region was removed from its collection bag and graded as to whether it was a primary guard hair (GH1), secondary guard hair (GH2), or underfur (UH). Figure 2A displays the scheme used to distinguish 20 primary from secondary guard hairs. Approximately 3-5 primary and secondary guard hairs were selected from each collection bag. Several hairs of each type were then plated onto their own glass microscope slides (Fisher Scientific) using Flotexx mounting medium (Lerner Cytology/Histology Reagents, Thermo Scientific - Model 137702). Flotexx was chosen because it has a refractive index (R.I.=1.52; 41) that is close to that of hair (R.I.=1.55; 3,7). Using mounting media with a refractive index that is close to that of the mounted object allows the examiner to obtain images that are free from shadow and contrast, which could otherwise mask the internal morphological characteristics of the hair in question (3,7). Each hair was oriented so that the basal region of the hair fiber corresponded with the left of the microscope slide. This served as an aid to the photographer to properly orient the hair under the microscope. Each hair on each slide was examined, and hairs were selected and photographed in a manner that documented microscopic fields that contained the most representative morphological characteristics for the particular hair type and section under view.

Preparation of Hair for Scanning Electron Microscopy Hair from the dorsal region was removed from its collection bag and graded as to whether it was a primary guard hair (GH1), secondary guard hair (GH2), or underhair (UH). Approximately five to ten primary guard hairs were removed to a sterile petri dish and dried in a desiccator for a period of 1-2 days. Scanning electron microscope mounts were then prepared using a sterile razor blade to excise a section of the hair’s shield region. The hair fiber was adhered to a metal specimen mount (Ted Pella, Inc. Redding, CA) with double stick carbon tape (Ted Pella, Inc. Redding, CA). The specimen and mount were then sputter-coated with gold at 6 mA voltage for 25 s. This procedure was performed once at 0 degrees 21

(specimen laying flat) and once at an angle of 45 degrees. Once the sputter-coating process was complete, the hair samples were viewed and photographed in a manner that documented microscopic fields that contained the most representative cuticular characteristics for the particular specimen and hair type under view.

Microscopy Slide and Stub Storage

Scanning Electron Microscope Specimen Mount Storage The gold-coated specimen mounts for Scanning Electron Microscopy were stored inside a desiccator in labeled cases (Ted Pella Inc.) for future use and reference.

Bright Field Microscopy Slide Storage Individual storage boxes (Fisher Scientific) were used for long-term storage of the bright field whole mount specimens. Boxes were separated by taxonomic order. Within the boxes, slides were organized alphabetically by genus. A legend was included with each slide storage box which provided information regarding genus, species, and individual body region corresponding to the numbered slot that the microscope slide occupies (see Fig. 8).

Digital Image Acquisition An Olympus BX45 transmitted light microscope coupled with a Q-imaging Retiga 2000R Camera was used to acquire digital images of the basal, sub-shield and shield portions of each hair specimen at 400 X magnification (Fig. 2C). Hair fibers too large for adequate viewing at 400 X magnification (e.g. the wild boar, Sus scrofa) were photographed at 200 X magnification. If variation was visualized along the shaft of the hair specimen, such as a marked increase or decrease in

22

FIG 8: Bright field microscopy slide storage. Microscope slides with whole mount specimens are on the left with a comprehensive listing of the slides on the right. diameter, medullary configuration change, or a change of the pigment color or density, then additional digital images were taken in order to record these variations. A Hitachi S-3500N Scanning Electron Microscope was used to obtain images of the cuticlar scale pattern of select specimens at 700 and 1,500 X magnification. The select specimens that were chosen represented the 21 families of the class Mammalia (1 representative from each family) that compose this database. This was done to show the wide variation regarding the cuticle pattern across family. Cuticle patterns were not recorded on a genus or species level, because, although differing characteristics may be observed, gross characteristics remain similar below the family taxonomic level (26). 23

A macroscopic and microscopic evaluation was conducted for each specimen. The classification scheme used, including cuticle scale descriptions, medullary configurations, and biological hair regions, was obtained from Moore and co-workers (5). The macroscopic characteristics recorded included hair color, form, and banding. Hair color was classified according to a Munsell color chart (42). Form refers to the overall look of the hair including shape and how that shape changes from the root to the tip. Banding patterns are an important classification in distinguishing mammals. Color banding is a band of color on a hair that is sharply separated from adjacent areas of different colors above and below the band (5,13,43). While many hairs are un-banded, the presence of banding is noteworthy, because this macroscopic feature can be used to distinguish certain taxa from others (5,13,34,43). Microscopic observations, such as medullary, corticullar, and cuticular characteristics, were then recorded according to a classification system adapted from Moore and co-workers (5). Medullary characteristics included a description and classification of the medulla type displayed. The shape of the cuticle scales and configuration is also described. Cortex characteristics such as pigment aggregation and color of pigment is also included in the evaluation. The classifications recorded and the classification system employed has been adapted from Moore and co-workers. Measurements of the medulla diameter, shaft diameter, and medullary index of the primary dorsal guard were carried out using Caseworks 2 software (Mideo Systems) after proper calibration with a scale micrometer (Fisher Scientific). 24 Photo Manipulation Adobe Photoshop version CS4 (Adobe Systems Incorporated) was used to prepare images for final publication. The image was cropped, placed on a uniform background, and the image properties (contrast, brightness, and color) were adjusted so that the internal characteristics of the hair could be best visualized in two dimensions. A photo template was created in Adobe Photoshop to optimize the process of image manipulation and final publication (see Fig. 9).

FIG 9: HAIRbase™ image template. The template was designed to aid in the consistent processing of multiple images of mammalian hair. Identifying numbers are in the upper left corner, scale for reference in the bottom left, and the file name of the specimen on the right side of the template.

25 Development of an Internet-based User Interface A user interface was created with iWeb ’09 version 2.0.4, a template-based website creation tool (Apple, Inc.), that allows the publishing of website content quickly and easily. The site is organized around a phylogenetic tree (44), which allows for easy navigation to each of the 16 orders of the class Mammalia that will be represented in the dataset. Individual order pages contain a list of species within that order that are contained within the database. Species pages display information regarding each individual specimen, including pelage observations, geographic distribution, and descriptions and measurements of dorsal guard hairs, photo-documentation of each hair specimen. Species pages generally reflect the variation present in a single individual. The images are separated into single albums displaying basal, sub-shield, and shield images of the primary and secondary guard hair from each of the particular areas on the specimen collected. The main infrastructure of the HAIRbase™ website has been constructed using the “Darkroom” template in iWeb ’09. Explanation for the creation of several types of webpages, including layout, formatting, and design features for the creation of order, species, image, and subspecies/breeds webpages using iWeb ’09 (see Appendix B).

RESULTS AND DISCUSSION

Development of HAIRbase

Rationale for Species Selection This work describes the creation of a database of primary and secondary guard hair images of domesticated, endangered, and non-protected mammals from around the world, which has been compiled largely through the use of bright field and scanning electron microscopy. HAIRbase™, as the database is known, covers variation across the class Mammalia by including a variety of mammals not previously seen in any atlas, including multiple regions on each specimen, multiple grades of hair, and multiple images of each hair. The dataset currently includes approximately 1,700 bright field microscopic images and 25 scanning electron microscopic images, with updates to be added frequently. HAIRbase™ is freely available on the Internet, allowing universal access and easy navigation through the collection. The implementation and the launching of HAIRbase™ on the Internet provides forensic investigators and researchers alike with a valuable resource for morphological comparison and animal identification. Contributions of additional or novel images from researchers may also be uploaded, with provenance data, making the collection interactive and expansive. HAIRbase™ was developed to include a representative collection of 133 species, which were chosen because of their significance to illegal wildlife trade (e.g., tiger, Panthera tigris; Tibetan antelope, Pantholops hodgsonii) or to facilitate legal trade in the fur industry (e.g., American mink, Mustela vison; American beaver, Castor canadensis). Many specimens were chosen as sole representatives of the family or genus in which they reside. The selected species come from the orders Artiodactyla and Carnivora, which comprise the majority of 27 the cases seen in forensic wildlife casework (45). Many of the selected species are listed as endangered or protected and are exceedingly rare in the wild, including 47 species that are protected under The Endangered Species Act of 1974 (see Appendix A,(15)) from the Artiodactyla and Carnivora orders of the class Mammalia. The current geographic focus for the mammals of HAIRbase is the mammals of North America, with select non-North American species that are seen in illegal trapping and poaching crimes, since the majority of forensic casework currently handled by the U.S. Fish and Wildlife Service involves furbearers and “trophy” animals. Additionally, of the 1,900 protected species of the class Mammalia, roughly 1,320 are native to North America (46). By having a North American focus, HAIRbase allows the inclusion of the greatest number of protected species while remaining centered on those species that seem to be most relevant to the greatest number of potential user groups. Several groups of mammals that are not endangered or trafficked are also included in HAIRbase as well. Although the database is arranged by species (the lowest taxonomic unit typically used in all organizational schemes of hair atlases), a comment is warranted here on the actual likelihood of making an identification to the species level based on hair microscopy. Claims of species-level identification are commonly attributed to microscopic hair identification (e.g., 47), but the reality is that genus- or family-level identifications are more likely (48). Forensic wildlife morphologist Bonnie C. Yates explains that hair is a conservative tissue and, therefore, differences in the structure of the hair do not typically change among congeners unless the function has changed evolutionarily (personal communication). These functional changes are usually seen at higher taxonomic 28 levels (i.e., genus or family). Therefore, identification at the genus or family level may be the best that can be attained with present scientific certainty.

Rationale for Classification of Morphological Characteristics Many classification schemes exist regarding the morphological structures of mammalian hair. The classification scheme employed by HAIRbase™ was presented by Thomas Moore and co-authors in 1974 (5). The classification schemes of the cellular organization of the medulla and cuticle can be seen in Figures 10 and 11, respectively. Individual hair fibers were classified using this scheme to correlate to the U.S. Fish and Wildlife Service that have also adopted this classification scheme for the use in wildlife forensic casework.

Rationale for Presentation of Micrographs Hair from the dorsal region of mammals has been studied and characterized for use in atlases and other academic references for many years (3,5,7,10-14,26). However, often an animal will display extreme variability on different areas of its body (21-22). To account for this variation, three areas on each specimen were collected and analyzed and have been included in HAIRbase™. These areas are the dorsal, ventral, and tail regions of the specimen. While other areas on the body can certainly be collected and displayed, these three regions were found to be the most variable between species, and so were chosen to represent the range of hair variation of the given mammals. Mammals have multiple grades of hair ranging from the long and coarse overhairs to the short and dense underhair or fur fibers (3,5,26). The transition of these grades of hair from one to another is more or less gradual; however, four grades of hair have been found to be distinguishable from one another (3). 29

FIG 10: Cellular classifications of the mammalian medulla. Reproduced with permission of author. Moore et al. 1974 (5). This figure shows medullary classifications that can be seen in a mammalian hair fiber.

30

FIG 11: Cuticular pattern classifications of mammalian hair. Reproduced with permission of author. Moore et al. 1974 (5). This figure shows cuticular classifications that can be seen in a mammalian hair fiber.

31

Underhair is much thinner and appears to have a wavy appearance (3,10, 26). Underhair remains relatively consistent in structure and function across the class Mammalia with few exceptions (3,10). Underhair was not chosen for inclusion in HAIRbase™, because this grade of hair is diagnostically unremarkable in isolation. Among overhairs, the primary guard hair is coarser, sometimes with a thickening in the distal region of the hair, known as the shield, and a thinner proximal region, known as the sub-shield and basal regions (3,5,7,10). The primary and secondary guard hairs were chosen for representation in HAIRbase™ because they display a number of characteristics that can be used for identification purposes (3,24). In addition to the variation of hair grades on each mammal, hairs can often display different characteristics depending on the linear position along the length of the hair being viewed (3,24,26). The three segments chosen for image capture – the basal, sub-shield, and shield regions of the hair – cover the widest range of variation along each hair shaft. The basal region of the hair is directly adjacent to the root. The sub-shield region is found on the shaft of the hair fiber. The shield is the area on the hair shaft that shows a marked increase in diameter. These areas were also chosen for inclusion in HAIRbase because they allow the viewing of a greater amount of characteristics and forensically are the areas that tend to be useful in hair comparisons (3,5,10).

Rationale for Website Presentation The HAIRbase™ website is the graphical user interface (GUI), which was designed to accommodate professionals with a thorough knowledge of hair classification and identification. The website is formed around a cladogram of the class Mammalia with links to the individual order webpages. A navigation bar 32 allows easy navigation to the “Welcome,” “Home,” “About,” “Browse,” “FBIC,” “NFWFL,” and “Contact” webpages. The “Home” webpage displays the central cladogram previously described. The “About” webpage describes the materials and methods that were used in the construction of HAIRbase™. The “Browse” page offers an additional way to browse the collection. This is done through a listing of the mammalian orders comprising the dataset with the number of species representatives that currently represent that particular order of mammal. The “FBIC” and “NFWFL” are links to the home pages of the Forensic Biotechnology Institute of California and the National Fish and Wildlife Forensic Laboratory, respectively. These two agencies collaborated to create HAIRbase™. The “Contact” link directs the user to contact information that will allow the contribution of verified hair samples to HAIRbase™. The individual order webpages contain information of the order as a whole (e.g., # of species, genera, and families included) and any identifying hair characteristics that are commonly used to separate the hair of animals of that particular order. The individual representatives of each of the orders are further separated to the family taxonomic level (e.g., Canidae, Felidae, etc.). This facilitates the easy navigation through the collection of species. Also, it has been found that closely related species exhibit similar morphological characteristics regarding their hair fibers (3,5,10). The organization and layout of the species page can be seen in its final form in Appendix C. The information included on these webpages was determined to be helpful for the successful determination of the origin of an unknown mammalian hair. An overall species image is included in the upper left corner of the page, adequately displaying the pelage and macroscopic appearance of the animal being viewed. Below the species image is a distribution map, displaying the geographical range of the given species as adapted from the IUCN Red List (49- 33

50). A link to a page presenting the scanning electron microscope images of an example of cuticular scalation at the familial level is included below the species distribution map. The right side of the webpage is dedicated to textual information including scientific classification of the animal (e.g., order, family, genus, species, etc.), conservation status, congeners to consider, pelage observations, and dorsal guard hair measurement and description. Conservation status refers to information regarding legal protection of the particular animal. The congeners of each species were also included, because the hair of closely related species is morphologically similar. This informs the viewer that these related mammals should be referred to in addition to the species illustrated for possible morphological similarity. Pelage observations refer to the macroscopic appearance of the animal (e.g., fur color variation, hair form, etc.). The dorsal guard hair measurement and description includes microscopic characterization of the guard hair depicted in the images. The measurements deemed appropriate for inclusion were: 1. Overall Length Measurement (in millimeters) 2. Shaft Diameter (in micrometers) 3. Medullary Index value (a proportion: medulla width/shaft width) (7) The images were organized into albums by body region, displaying images of 3 morphological regions on each hair (basal, subshield, and shield). If these body regions were not available, another body region was chosen to display the morphological variation within the specimen. The individual images were organized into columns, with the primary guard hair images on the left and secondary guard hair images on the right. The basal, subshield, and shield regions were organized so that the user can easily view the morphological characteristics of the different regions of the hair shaft, as well as perform a cross comparison between the primary and secondary guard hairs. Provenance information, total 34 magnification, and any notes that may be of particular interest to the viewer or explanatory in nature (e.g., presence of clearing in the medulla) were included with the images. With the layout described above, the combination of ease of use and the information that the site contains will serve as a valuable applicative tool to professional trace evidence examiners.

User Guidelines The arena of endangered species law and trade statutes are constantly changing and being updated (51). Since HAIRbase will remain “active” on the Internet, with more specimens, information, and images constantly being added, the information given about the conservation status for each species can change along with these regulations. The importance of a reliable online reference that can adapt to the changing needs of its user groups ensures that HAIRbase™ will remain a relevant reference for forensic professionals. Almost as integral as the information contained within the database is the proper utilization of such a tool. The effective use of HAIRbase™, while begun from a perspective of wildlife forensic trace evidence analysis, remains similar in application across multiple disciplines dealing with hair identification. HAIRbase™ is intended to show variability and subtleties of animal hair structure and function. It is not intended as a replacement for microscopical comparison of actual animal hairs. Morphology, as a comparative science, ultimately relies on the use of a reference standard, that is, an actual hair from an identified source. The images collected in this database should not, nor were they meant to, be used alone or exclusively for an identification. The user is encouraged to always compare actual hair standards from known specimens for direct comparisons, whether microscopically or macroscopically. 35

The importance of reliable hair standards for hair identification is unquestioned. However, references and atlases play a large role in prompting further investigation of certain species and specimens. Atlases can also function as a type of initial screening process before preparing whole mounts of specimens and comparing to actual standards from a reference collection. HAIRbase™ is specifically geared to the forensic community, and is set apart because it approaches the study of hair morphology by stressing the concept of variation. Hair is commonly seen in forensic casework and analyzed in a standardized manner reliant on comparison with a prepared reference sample. However, before this microscopical comparison can take place, the forensic analyst must generate investigative leads by eliminating certain species and considering ones that show similarities to the unknown sample. HAIRbase™ has been designed and constructed to be a launching point for further investigation, with the atlas implemented as a type of screening tool to eliminate unlikely species and further investigate similar candidates. Through the implementation of HAIRbase™, the time and resources required for the initiation of a hair examination are dramatically reduced.

Future Research A great strength of HAIRbase™, and of dissemination on the Internet in general, is the ease of integration of new information and new techniques. The initial design and infrastructure of HAIRbase™ has been described here. However, because the website remains live on the Internet, there are multiple scientific avenues that can and should be pursued to ensure the professional use and application of HAIRbase™. 36

Internet dissemination allows the inclusion and analysis of more species within the class Mammalia quickly and easily. Containing approximately 5,400 species, the class Mammalia is extremely large and equally diverse. This thesis has described the analysis of 133 such mammals belonging to the Artiodactyla and Carnivora orders. However, there are other orders that remain undone and are forensically relevant. One forensically relevant order that requires classification and analysis is Primates. The development of a domestic section will also aid forensic examiners to aid in civil litigations. The inclusion of these species will further aid forensic professionals by giving them a reference for the variation occurring in non-human primates (i.e., monkeys and apes) and domestic species (i.e., dog, cats, sheep, etc.). Owing to the overall coverage of variation among and within species, more body regions on each specimen can be collected and analyzed. More images of each hair fiber can be included in the dataset, as well as different grades of hair (other than primary and secondary guard hairs) on the same specimen. The cuticle scale pattern images can be expanded to the genera or species level. Currently, cuticle scale SEM images were taken at the family taxonomic level. Also, more images of the cuticle scale pattern can be captured. New techniques can certainly be applied to the hair fibers under study. These techniques include taking transverse and longitudinal cross-sections of the hair fibers. The viewing of transverse cross-sections allows the analysis of the cross-sectional shape of the hair, which can be used to separate certain species (10). Longitudinal cross-sections permits “clearing” of the medulla, by allowing the infiltration of mounting media into the medullar cells and driving out the trapped air. This technique gives a clear view of the cellular composition of the medulla and can aid in hair identification by allowing the easy viewing of 37 morphological characteristics that may be obscured by dense pigment, or air in the medulla (24). This discussion has been concerned with the morphological features of the hair of these selected mammals. However, there is no reason why HAIRbase™ cannot expand into the genetic realm. That is why HAIRbase™ is infinitely expandable. With multiple scientific avenues to consider, HAIRbase™ can truly become a valuable resource for forensic trace examiners and beyond.

SUMMARY

HAIRbase is a flexible online atlas of hair morphology, specifically geared to the needs of the wildlife forensic community, that attempts to address the deficiencies of past atlases by providing information on the characteristics of a given species as a whole while, at the same time, providing enough coverage of a given individual to adequately represent the amount of variation in the grades of hair that are generally encountered over an individual’s entire body. HAIRbase currently includes 133 species chosen to represent the variation in the Artiodactyla and Carnivora orders within the class Mammalia, and for many species will include two grades of hair from three regions on the body of each individual, as well as three positional images from each hair. The user interface of HAIRbase is designed to allow easy navigation through the collection. Individual species’ web pages display information regarding the specimen including observations of pelage, geographic distribution, and dorsal guard hair description and measurement, as well as the images captured for that particular specimen. The images are separated into individual albums displaying basal, sub-shield, and shield images of the primary and secondary guard hair from each of the particular areas on the specimen collected. The great strength of this database is the capability to add more specimens, allowing the database to grow and further accommodate the needs of the forensic trace examiner, as well as other professionals in scientific disciplines that employ morphology in the identification of trace material.

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APPENDICES

APPENDIX A: SPECIES INCLUDED IN HAIRBASE™

Order Family Genus Species Common Animal Name Conservation Status Artiodactyla Antilocapridae Antilocapra americana Pronghorn Antelope least Bovidae Aepyceros melampus Impala least Alcelaphus buselaphus Kongoni Hartebeest least Antidorcas marsupialis Springbuck least Bison bison Bison near threatened Bos grunniens Yak vulnerable Javanicus Java Banteng endangered primigenius Cow extinct in the wild taurus Domesticated Cow domestic Bubalis mindorensis Tamaraw critically endangered Budorcas taxicolor Sichuan Takin vulnerable Capra caucasian West caucasian Tahr endangered hircus Goat domestic ibex Alpine Ibex least Capricornus sumatraensis Mainland Serow near threatened Cephalophus zebra Zebra Duiker vulnerable Cervus elaphus Red Deer least Connochaetes taurinus Bridled gnu-wildebeest least 46

Gazella rufifrons Red-fronted Gazelle vulnerable Hippotragus equinus Roan Antelope least niger Sable Antelope least Kobus ellipsiprymnus Waterbuck least Litocranius walleri Gerenuk near threatened Mardoqua sp Dik Dik least Muntiacus reevesi Chinese Muntjac least Naemorhedus goral Himalayan Goral near threatened Nanger granti Grant's Gazelle least Nilgitragus hylocrius Nilgiri Tahr endangered Oreamnos americanus Mountain Goat least Oreotragus oreotragus Klipspringer least Oryx beisa Beisa Oryx near threatened gazella Gemsbok least Ovibus moschatus Muskox least Ovis ammon Mountain Sheep near threatened canadensis Mountain Big Horn Sheep least dalli White Sheep least dalli Dall Sheep least domestica Lincoln Cross Sheep domestic ammon polii Marco Polo Sheep near threatened Pantholops hodgsonii Tibetan Antelope endangered Philantomba monticola Blue Duiker least Pseudois nayaur Chinese Bharal least Raphicerus campestris Steinbuck least Redunca redunca Bohor Reedbuck least 47

Rupicapra rupicapra Chamois least Saiga tatarica Saiga Antelope critically endangered Sylvicapra grimmia Crowned Duiker least Taurotragus derbianus Giant Eland least Tragelaphus strepsiceros Greater Kudu least Tragelaphus anagasii Lowland Nyala least Camelidae Alces alces Moose least Axis porcinus Hog Deer endangered Camelus taur Dromedary Camel domestic Capriolus capriolus Roe Deer least Cervidae Cervus nippon Sika Deer least canadensis Elk least Dama dama Fallow Deer least Elaphurus davidianus Pere David's Deer extinct in the wild Lama glama Llama domestic guanicoe Guanaco least Moschus mosciferus Siberian Musk Deer vulnerable Muntiacus reevesi Reeve's Muntjac least Odocoileus hemionus Mule Deer least virginianus White-tailed Deer least Rangifer articus Barren Ground Carribou least tarandus Caribou least Vicugna vicugna Vicugna least Giraffidae Giraffa camelopardalis Giraffe least Okapi johnstonii Okapi near threatened Suidae Sus domestica Pig domestic 48

scrofa Wild Boar least Tayassuidae Pecari tajacu Collared Peccarry least Tragulidae Tragulus jovanicus Lesser Mouse Deer least Carnivora Ailuridae Ailurus fulgens Red Panda vulnerable Canidae Canis aureus Golden Jackal least latrans Coyote least lupus Grey Wolf least lupus domesticus Dog domestic Cuon alpinus Dhole endangered Lycaon pictus African Wild Dog endangered Nyctereutes procyonoides Raccoon Dog least Urocyon cinereoargenteus Grey Fox least Vulpes vulpes Red Fox least Felidae Acinonyx jubatus Cheetah vulnerable Felis catus Cat domestic domestica Domesticated Cat domestic Leopardus pardalis Ocelot least Linx canadensis Lynx least Lynx rufus Bobcat least Neofelis nebulosa Clouded Leopard vulnerable Panthera leo Lion vulnerable Onca Jaguar near threatened pardus Leopard near threatened tigris Tiger vulnerable tigris sumatrae Sumatran Tiger critically endangered tigris tigris Bengal Tiger endangered 49

Prionailurus bengalensis Leopard Cat least Puma concolor Puma least yaguarundi Jaguarundi least Uncia uncia Snow Leopard endangered Herpestidae Suricata suricata African Meerkat least Hyaenidae Crocuta crocuta Spotted Hyaena least Hyaena brunnea Brown Hyaena near threatened Mephitidae Mephitis mephitis Striped Skunk least Spilogale putorius Eastern Spotted Skunk least Mustelidae Aonyx cinerea Oriental Small-clawed Otter vulnerable Enhydra lutris Sea Otter endangered Gulo gulo Wolverine least Lontra canadensis North American River Otter least Martes caurina American Marten least zibellina Sable least Mustela erminea Ermine least frenata Long-tailed Weasel least nigripes Black-footed Ferret endangered putorius Ferret domestic siberica Siberian Mink least vison American Mink least Taxidea taxus American Badger least Martes pennanti Fisher least Otariidae Calorhinus ursinus Northern Fur Seal vulnerable Zalophus californicus Sea Lion least Phocidae Phoca vitulina Harbor Seal least 50

Procyonidae Bassariscus astutus Ringtail Cat least Nasua narica White-nosed Coati least Procyon lotor Raccoon least Potos flavus Kinkajou least Ursidae Ailuropoda melanoleuca Giant Panda endangered Genetta thierryi Genet Cat least Ursus arctos Brown Bear least Helarctos malayanus Malayan Sunbear vulnerable Melursus ursinus Sloth Bear vulnerable Tremarctos ornatus Sloth Bear vulnerable Ursus americanus Black Bear least thibetanus Asiatic Black Bear vulnerable maritimus Polar Bear vulnerable arctos horribilis Grizzly Bear vulnerable

Bolded species are currently protected under the Endangered Species act of 1974.

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APPENDIX B: WEBPAGE TEMPLATES

53

Order webpage template 54

Species webpage template 55

Album webpage template 56

Subspecies/Breed webpage template

APPENDIX C: HAIRBASE™ IDENTIFICATION NUMBERS AND CONGENERS

Order Common Name Identification Congeners Number Artiodactyla Pronghorn Antelope BA516 None Addax NFWFL Coll. None Impala NFWFL Coll. None Kongoni Hartebeest CSU BGC 01 caama, lichtensteinii Springbok CSU BGC 02 None American Bison NFWFL Coll. bonasus Yak NFWFL Coll. frontalis, javanicus, sauveli, taurus Java Banteng NFWFL Coll. frontalis, grunniens, sauveli, taurus Domestic Cow NFWFL Coll. frontalis, grunniens, javanicus, sauveli Tamaraw NFWFL Coll. bubalis, depressicornis, quarlesi Sichuan Takin NFWFL Coll. None West Caucasian Tur NFWFL Coll. falconeri, hircus, ibex, nubiana, pyrenaica, sibirica, walie Goat NFWFL Coll. caucasica, falconeri, ibex, nubiana, pyrenaica, sibirica, walie Alpine Ibex NFWFL Coll. caucasica, falconeri, hircus, nubiana, pyrenaica, sibirica, walie Mainland Serow NFWFL Coll. crispus, milneedwardsii, rubidus, swinhoei, thar Zebra Duiker NFWFL Coll. adersi, brookei, callipygus, dorsalis, jentinki, leucogaster, natalensis, niger, nigrifrons, ogilbyi, rufilatus, silvicultor, spadix, weynsi Blue Wildebeest CSU BGC 04 gnou Red-fronted Gazelle CSU BGC 06 rufina, thomsonii Nilgiri Tahr NFWFL Coll. jayakari, jemlahicus 58

Roan Antelope NFWFL Coll. leucophaeus, niger Sable Antelope NFWFL Coll. equinus, leucophaeus Waterbuck CSU BGC 07 kob, leche, megaceros, vardonii Gerenuk CSU BGC 08 None Dik Dik CSU BGC 09 guentheri, kirkii, piacentinii, saltiana Himalayan Goral NFWFL Coll. baileyi, caudatus, griseus Grant's Gazelle CSU BGC 10 dama, soemmerringii Mountain Goat NFWFL Coll. None Klipspringer CSU BGC 11 None Beisa Oryx CSU BGC 12 dammah, gazella, leucoryx Gemsbok NFWFL Coll. beisa, dammah, leucoryx Muskox NFWFL Coll. None Argali NFWFL Coll. aries, canadensis, dalli, nivicola Marco Polo Sheep NFWFL Coll. aries, canadensis, dalli, nivicola Lincolnshire Cross mam 612 ammon, canadensis, dalli, nivicola Sheep Mountain Big Horn NFWFL Coll. ammon, aries, dalli, nivicola Sheep Dall's Sheep NFWFL Coll. ammon, aries, canadensis, nivicola Chiru NFWFL Coll. None Blue Duiker NFWFL Coll. maxwellii Chinese Bharal NFWFL Coll. schaeferi Steenbok CSU BGC 14 melanotis, sharpei Bohor Reedbuck CSU BGC 15 arundinum, fulvorufula Chamois NFWFL Coll. pyrenaica Saiga Antelope NFWFL Coll. borealis 59

Common Duiker NFWFL Coll. None Cape Buffalo CSU BGC 16 None Giant Eland NFWFL Coll. oryx Lowland Nyala NFWFL Coll. buxtoni, eurycerus, imberbis, scriptus, spekii, strepsiceros Greater Kudu CSU BGC 17 angasii, buxtoni, eurycerus, imberbis, scriptus, spekii Dromedary Camel NFWFL Coll. bactrianus, dromedarius Llama NFWFL Coll. None Guanaco NFWFL Coll. None Vicugna NFWFL Coll. None Moose NFWFL Coll. americanus Hog Deer NFWFL Coll. axis, calamianensis, kuhlii Roe Deer NFWFL Coll. pygargus Elk CSU BGC 03 elaphus, nippon Red Deer NFWFL Coll. nippon Sika Deer NFWFL Coll. elaphus Fallow Deer N1304 None Pere David's Deer NFWFL Coll. None Siberian Musk Deer 39637 anhuiensis, berezovskii, chrysogaster, cupreus, fuscus, leucogaster Indian Muntjac NFWFL Coll. atherodes, crinifrons, feae, gongshanensis, puhoatensis, putaoensis, reevesi, rooseveltorum, truongsonensis, vuquangensis Reeve's Muntjac NFWFL Coll. atherodes, crinifrons, feae, gongshanensis, muntjak, puhoatensis, putaoensis, rooseveltorum, truongsonensis, vuquangensis Mule Deer mam 214 virginianus White-tailed Deer NFWFL Coll. hemionus Caribou NFWFL Coll. None 60

Giraffe NFWFL Coll. None Okapi 35928 None Pig NFWFL Coll. ahoenobarbus, barbatus, bucculentus, cebifrons, celebensis, oliveri, philippensis, salvanius, verrucosus Wild Boar mam 1718 ahoenobarbus, barbatus, bucculentus, cebifrons, celebensis, oliveri, philippensis, salvanius, verrucosus Collared Peccary NFWFL Coll. None Lesser Mouse Deer NFWFL Coll. kanchil, napu, nigricans, versicolor, williamsoni Carnivora Red Panda NFWFL Coll. None Golden Jackal NFWFL Coll. adustus, latrans, lupus, mesomelas, simensis Dog NFWFL Coll. adustus, aureus, latrans, lupus, mesomelas, simensis Coyote N1115 adustus, aureus, lupus, mesomelas, simensis Grey Wolf N1081 adustus, aureus, latrans, mesomelas, simensis Dhole 501049 None African Wild Dog NFWFL Coll. None Raccoon Dog NFWFL Coll. None Grey Fox CSU 1506 littoralis Red Fox N1087 lagopus, macrotis, pallida, bengalensis, cana, chama, corsac, ferrilata, rueppelii, velox, zerda Cheetah NFWFL Coll. None Domestic Cat NFWFL Coll. bieti, catus, chaus, manul, margarita, nigripes, silvestris Ocelot NFWFL Coll. braccatus, colocolo, geoffroyi, guigna, jacobitius, pajeros, tigrinus, wiedii Lynx N1245 lynx, pardinus, rufus Bobcat CSU 1705 canadensis, lynx, pardinus Clouded Leopard NFWFL Coll. None Lion NFWFL Coll. onca, pardus, tigris 61

Jaguar NFWFL Coll. leo, pardus, tigris Leopard NFWFL Coll. leo, onca, tigris Sumatran Tiger MO 608344 leo, onca, pardus Bengal Tiger 591444 leo onca, pardus Leopard Cat NFWFL Coll. iriomotensis, planiceps, rubiginosus, viverrinus Puma N1240 yagouaroundi Jaguarundi NFWFL Coll. concolor Snow Leopard NFWFL Coll. None African Meerkat 35944 None Spotted Hyena NFWFL Coll. None Brown Hyena 789503, 1449 hyaena Striped Skunk NFWFL Coll. macroura Eastern Spotted NFWFL Coll. aquaticus, angustifrons, gracilis, pygmaea Skunk Oriental small NFWFL Coll. capensis clawed otter Sea Otter NFWFL Coll. None Wolverine NFWFL Coll. None North American mam 075 felina, longicaudis, provocax Otter American Marten NFWFL Coll. americana, flavigula, foina, gwatkinsii, martes, melampus, pennanti, zibellina Fisher NFWFL Coll. americana, flavigula, foina, gwatkinsii, martes, melampus, zibellina Sable NFWFL Coll. americana, flavigula, foina, gwatkinsii, martes, melampus, pennanti Ermine CSU 69 eversmanii, africana, altaica, felipei, frenata, itatsi, kathiah, 62

lutreola, lutreolina, nigripes, nivalis, nudipes, putorius, sibirica, strigidorsa, subpalmata Long-tailed Weasel CSU 74 erminea, eversmanii, africana, altaica, felipei, itatsi, kathiah, lutreola, lutreolina, nigripes, nivalis, nudipes, putorius, sibirica, strigidorsa, subpalmata Black-footed Ferret NFWFL Coll. erminea, eversmanii, africana, altaica, felipei, frenata, itatsi, kathiah, lutreola, lutreolina, nivalis, nudipes, putorius, sibirica, strigidorsa, subpalmata Ferret NFWFL Coll. erminea, eversmanii, africana, altaica, felipei, frenata, itatsi, kathiah, lutreola, lutreolina, nigripes, nivalis, nudipes, sibirica, strigidorsa, subpalmata Siberian Mink NFWFL Coll. erminea, eversmanii, africana, altaica, felipei, frenata, itatsi, kathiah, lutreola, lutreolina, nigripes, nivalis, nudipes, strigidorsa, subpalmata American Mink CSU 2818 macrodon American Badger NFWFL Coll. None Northern Fur Seal NFWFL Coll. None Sea Lion NFWFL Coll. japonicus, wollebaeki Harbor Seal NFWFL Coll. largha Ringtail Cat NFWFL Coll. sumichrasti White-nosed Coati NFWFL Coll. nasua Kinkajou mam 1343 None Raccoon CSU 1508 cancrivorus, pygmaeus Giant Panda NFWFL Coll. None Malayan Sunbear NFWFL Coll. None Sloth Bear NFWFL Coll. None Spectacled Bear NFWFL Coll. None 63

American Black N1125 arctos, maritimus, thibetanus Bear Brown Bear NFWFL Coll. americanus, maritimus, thibetanus Grizzly Bear CSU BGC 18 americanus, arctos, maritimus, thibetanus Polar Bear NFWFL Coll. americanus, arctos, thibetanus Asiatic Black Bear NFWFL Coll. americanus, arctos, maritimus Genet Cat NFWFL Coll. abyssinica, angolensis, bourloni, cristata, genetta, johnstoni, maculata, pardina, piscivora, poensis, servalina, tigrina, victoriae

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APPENDIX D: HAIRBASE™ SPECIES COLLECTION AND STORAGE CHECKLIST

Order Common Backer Small Large Dorsal Ventral Tip of Tail Other Regions Name Page Labels Labels Collected and Collected and Collected and Collected and Stored Stored Stored Stored Artiodactyla Pronghorn 06/15/10 06/21/10 07/12/10 06/30/10 06/30/10 None Available Forehead Antelope Addax 06/14/10 02/08/10 07/12/10 06/30/10 06/30/10 06/30/10 None Impala 06/14/10 02/08/10 07/12/10 06/30/10 06/30/10 None Available Forehead Kongoni 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead Hartebeest Springbok 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead American 06/14/10 02/08/10 07/12/10 06/30/10 06/30/10 06/30/10 None Bison Yak 06/14/10 02/08/10 07/12/10 06/30/10 06/30/10 06/30/10 None Java Banteng 06/14/10 02/08/10 02/08/10 06/28/10 06/28/10 06/28/10 Forehead, Top of Foot, back of thigh, vibrissae Domestic 06/14/10 03/15/10 07/12/10 06/30/10 06/30/10 06/30/10 None Cow Tamaraw 06/15/10 02/09/10 07/12/10 06/30/10 06/30/10 None Available Forehead Sichuan 06/15/10 02/09/10 02/09/10 06/28/10 06/28/10 06/28/10 Forehead, Top of Foot, Takin back of thigh, vibrissae West 06/15/10 02/09/10 07/12/10 06/30/10 06/30/10 06/30/10 None Caucasian Tur Goat 06/15/10 02/09/10 07/12/10 07/01/10 07/01/10 07/01/10 None Alpine Ibex 06/15/10 02/08/10 07/12/10 06/30/10 06/30/10 06/30/10 None Mainland 06/15/10 02/08/10 07/12/10 06/30/10 06/30/10 None Available Forehead Serow Zebra Duiker 06/07/10 02/09/10 02/09/10 06/28/10 06/28/10 06/28/10 Forehead, Top of Foot, back of thigh, vibrissae Blue 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead Wildebeest Red-fronted 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead 66

Gazelle Nilgiri Tahr 06/07/10 02/09/10 07/12/10 07/01/10 07/01/10 07/01/10 None Roan 06/07/10 07/15/10 07/12/10 06/30/10 06/30/10 06/30/10 Forehead, Back of Antelope Thigh Sable 06/07/10 06/30/10 07/12/10 06/30/10 06/30/10 06/30/10 None Antelope Waterbuck 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead Gerenuk 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead Dik Dik 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead Himalayan 06/18/10 02/09/10 07/12/10 06/30/10 06/30/10 06/30/10 None Goral Grant's 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead Gazelle Mountain 06/18/10 02/09/10 07/12/10 07/01/10 07/01/10 07/01/10 None Goat Klipspringer 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead Beisa Oryx 06/18/10 06/21/10 07/12/10 06/21/10 06/21/10 None Available Forehead Gemsbok 06/18/10 02/09/10 07/12/10 07/01/10 07/01/10 07/01/10 None Muskox 06/18/10 02/09/10 07/12/10 07/01/10 07/01/10 07/01/10 None Argali 06/07/10 02/09/10 07/12/10 07/01/10 None Available None Available Top of Foot Marco Polo 06/18/10 02/09/10 07/12/10 07/01/10 07/01/10 07/01/10 None Sheep Lincolnshire 06/07/10 02/09/10 02/09/10 06/28/10 06/28/10 06/28/10 Forehead, Top of foot, Cross Sheep Back of Thigh, Vibrissae Mountain Big 06/07/10 02/09/10 07/12/10 07/01/10 07/01/10 07/01/10 None Horn Sheep Dall's Sheep 06/18/10 02/09/10 07/15/10 07/01/10 07/01/10 07/01/10 None Chiru 06/18/10 02/09/10 07/15/10 07/01/10 07/01/10 None Available Forehead Blue Duiker 06/18/10 02/10/10 07/15/10 07/01/10 07/01/10 07/01/10 None 67

Chinese 06/18/10 02/10/10 07/15/10 07/01/10 07/01/10 07/01/10 None Bharal Steenbok 06/18/10 06/21/10 07/15/10 06/21/10 06/21/10 None Available Forehead Bohor 06/18/10 06/21/10 07/15/10 06/21/10 06/21/10 None Available Forehead Reedbuck Chamois 06/07/10 02/10/10 07/15/10 07/01/10 07/01/10 None Available Forehead Saiga 06/07/10 02/10/10 06/30/10 06/26/10 06/26/10 06/26/10 None Antelope Common 06/07/10 02/10/10 07/15/10 07/01/10 07/01/10 07/01/10 None Duiker Cape Buffalo 06/18/10 08/03/10 07/15/10 06/21/10 06/21/10 None Available Forehead Giant Eland 06/07/10 02/10/10 02/10/10 06/28/10 06/28/10 06/28/10 Forehead, Top of Foot, back of thigh, vibrissae Lowland 06/07/10 02/10/10 02/10/10 06/28/10 06/28/10 06/28/10 Top of foot, Back of Nyala Thigh, Forehead Greater Kudu 06/18/10 06/21/10 07/15/10 06/21/10 06/21/10 None Available Forehead Dromedary 06/08/10 02/10/10 07/15/10 06/30/10 06/30/10 06/30/10 None Camel Llama 06/08/10 02/10/10 07/15/10 06/30/10 06/30/10 06/30/10 None Guanaco 06/08/10 02/10/10 07/15/10 06/30/10 06/30/10 06/30/10 None Vicugna 06/08/10 02/10/10 07/15/10 07/01/10 07/01/10 07/01/10 None Moose 06/08/10 06/21/10 07/15/10 06/30/10 None Available 06/30/10 Forehead Hog Deer 06/08/10 02/10/10 07/15/10 06/30/10 06/30/10 06/30/10 None Roe Deer 06/08/10 02/10/10 02/10/10 06/28/10 06/28/10 06/28/10 Forehead, Top of Foot, back of thigh, vibrissae Elk 06/18/10 06/21/10 07/15/10 06/21/10 06/21/10 None Available Forehead Red Deer 06/15/10 02/10/10 07/15/10 06/30/10 06/30/10 06/30/10 None Sika Deer 06/15/10 02/10/10 07/15/10 06/30/10 06/30/10 06/30/10 None Fallow Deer 06/15/10 09/04/10 07/15/10 06/30/10 06/30/10 None Available Forehead, Vibrissae Pere David's 06/15/10 02/10/10 02/10/10 06/28/10 06/28/10 06/28/10 Forehead, Top of Foot, 68

Deer back of thigh, vibrissae Siberian 06/15/10 02/10/10 07/15/10 07/01/10 06/30/10 06/30/10 None Musk Deer Indian 06/15/10 02/11/10 06/26/10 06/26/10 06/26/10 06/26/10 Forehead, Top of Foot, Muntjac back of thigh, vibrissae Reeve's 06/15/10 02/11/10 02/11/10 06/28/10 06/28/10 06/28/10 Forehead, Top of Foot, Muntjac back of thigh, vibrissae Mule Deer 06/15/10 02/11/10 07/15/10 07/01/10 07/01/10 07/01/10 None White-tailed 06/15/10 02/11/10 07/15/10 06/26/10 06/26/10 06/26/10 None Deer Caribou 06/11/10 02/11/10 06/30/10 06/26/10 06/26/10 06/26/10 None Giraffe 06/15/10 06/21/10 07/15/10 06/30/10 06/30/10 06/30/10 None Okapi 06/15/10 02/11/10 07/15/10 07/01/10 07/01/10 07/01/10 Back of Thigh, Forehead Pig 06/15/10 02/11/10 02/11/10 07/01/10 07/01/10 07/01/10 None Wild Boar 06/15/10 02/11/10 07/15/10 07/01/10 07/01/10 07/01/10 None Collared 06/15/10 02/11/10 07/15/10 07/01/10 07/01/10 07/01/10 Unspecified Peccary Lesser Mouse 06/11/10 02/11/10 07/15/10 07/01/10 07/01/10 07/01/10 None Deer Carnivora Red Panda 06/11/10 02/11/10 06/30/10 06/23/10 06/23/10 06/23/10 None Golden 06/11/10 02/11/10 06/30/10 06/23/10 06/23/10 06/23/10 None Jackal Dog 06/11/10 02/11/10 02/11/10 07/01/10 07/01/10 07/01/10 None Coyote 06/09/10 02/11/10 02/11/10 06/28/10 06/28/10 06/28/10 Forehead, Top of foot, Back of Thigh, Vibrissae Grey Wolf 06/11/10 02/11/10 02/11/10 06/28/10 06/28/10 06/28/10 Forehead, Top of foot, Back of Thigh, Vibrissae Dhole 06/09/10 02/11/10 06/30/10 07/15/10 06/28/10 06/28/10 None 69

African Wild 06/09/10 02/12/10 02/12/10 06/28/10 06/28/10 06/28/10 Forehead, Top of foot, Dog Back of Thigh, Vibrissae Raccoon Dog 06/09/10 02/12/10 06/30/10 06/23/10 06/23/10 06/23/10 None Grey Fox 06/15/10 12/03/10 12/03/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh Red Fox 06/09/10 02/12/10 02/12/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh Cheetah 06/11/10 09/04/10 06/30/10 06/23/10 06/23/10 06/23/10 None Domestic Cat 06/11/10 02/12/10 02/12/10 07/01/10 07/01/10 07/01/10 None Ocelot 06/11/10 02/12/10 06/30/10 06/23/10 06/23/10 06/23/10 None Lynx 06/11/10 02/12/10 06/30/10 06/23/10 06/23/10 06/23/10 None Bobcat 06/15/10 02/12/10 02/12/10 06/28/10 06/28/10 06/28/10 Forehead, Top of Foot, back of thigh, vibrissae Clouded 06/11/10 02/12/10 06/30/10 06/23/10 06/23/10 06/23/10 None Leopard Lion 06/18/10 09/04/10 06/30/10 06/23/10 06/23/10 06/23/10 None Jaguar 06/18/10 02/12/10 06/30/10 06/23/10 06/23/10 06/23/10 None Leopard 06/18/10 09/04/10 06/30/10 06/23/10 06/23/10 06/23/10 None Sumatran 06/18/10 02/12/10 06/30/10 06/23/10 06/23/10 06/23/10 None Tiger Bengal Tiger 06/15/10 02/12/10 06/30/10 06/23/10 06/23/10 06/23/10 None Leopard Cat 06/15/10 02/12/10 06/30/10 06/23/10 06/23/10 None Available Forehead Puma 06/15/10 02/12/10 06/30/10 06/23/10 06/23/10 06/23/10 None Jaguarundi 06/15/10 02/12/10 02/12/10 06/28/10 06/28/10 06/28/10 Forehead, Top of Foot, back of thigh, vibrissae Snow 06/11/10 02/15/10 06/30/10 06/23/10 06/23/10 06/23/10 None Leopard African 06/11/10 02/15/10 07/15/10 None Available 06/23/10 06/23/10 Forehead 70

Meerkat Spotted 06/11/10 02/15/10 07/15/10 06/23/10 06/23/10 06/23/10 None Hyena Brown Hyena 06/18/10 02/15/10 07/15/10 06/23/10 06/23/10 06/23/10 None Striped 06/18/10 02/15/10 06/23/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Skunk Vibrissae, Back of thigh Eastern 06/11/10 02/15/10 07/15/10 06/23/10 06/23/10 06/23/10 None Spotted Skunk Oriental 06/11/10 02/15/10 07/15/10 06/23/10 06/23/10 06/23/10 None small clawed otter Sea Otter 06/09/10 02/15/10 07/15/10 06/23/10 06/23/10 06/23/10 None Wolverine 06/09/10 02/15/10 07/15/10 06/28/10 06/28/10 06/28/10 None North 06/09/10 02/15/10 07/15/10 06/23/10 06/23/10 06/23/10 None American Otter American 06/09/10 02/15/10 06/23/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Marten Vibrissae, Back of thigh Fisher 06/15/10 02/15/10 06/23/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh Sable 06/15/10 02/15/10 07/15/10 06/23/10 06/23/10 06/23/10 None Ermine 06/15/10 02/15/10 06/23/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh Long-tailed 06/15/10 02/15/10 06/23/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Weasel Vibrissae, Back of thigh Black-footed 06/15/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Ferret 71

Ferret 06/15/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Siberian 06/08/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Mink American 06/08/10 02/16/10 02/16/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Mink Vibrissae, Back of thigh American 06/08/10 02/16/10 02/16/10 06/28/10 06/28/10 06/28/10 Forehead, Top of foot, Badger Back of Thigh, Vibrissae Northern Fur 06/08/10 02/16/10 07/15/10 06/23/10 06/23/10 None Available Forehead Seal Sea Lion 06/08/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Harbor Seal 06/07/10 02/16/10 07/15/10 06/23/10 06/23/10 None Available Forehead Ringtail Cat 06/07/10 02/16/10 02/16/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh White-nosed 06/07/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Coati Kinkajou 06/07/10 02/16/10 02/16/10 06/28/10 06/28/10 06/28/10 Forehead, Top of foot Raccoon 06/07/10 02/16/10 06/23/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh Giant Panda 06/07/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Malayan 06/07/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Sunbear Sloth Bear 06/07/10 02/16/10 06/23/10 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh Spectacled 06/07/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Bear American 06/07/10 02/16/10 02/16/10 06/28/10 06/28/10 06/23/10 Top of foot, Forehead, Black Bear Vibrissae, Back of thigh 72

Brown Bear 06/07/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Grizzly Bear 06/18/10 06/21/10 07/15/10 06/21/10 06/21/10 06/21/10 None Polar Bear 06/07/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Asiatic Black 06/07/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None Bear Genet Cat 06/07/10 02/16/10 07/15/10 06/23/10 06/23/10 06/23/10 None

73

APPENDIX E: HAIRBASE™ MICROSCOPE PREPARATION CHECKLIST

Order Common Name Dorsal Plated Ventral Plated Tip of Tail Plated Others Body Parts Plated Artiodactyla Pronghorn Antelope 06/23/10 06/23/10 None Available Forehead Addax 06/15/10 06/15/10 06/15/10 None Available Impala 06/23/10 06/23/10 None Available Forehead Kongoni Hartebeest 06/23/10 06/23/10 None Available Forehead Springbok 06/23/10 06/23/10 None Available Forehead American Bison 06/23/10 06/23/10 06/23/10 None Available Yak 06/15/10 06/15/10 06/15/10 None Available Java Banteng 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Domestic Cow 06/15/10 06/15/10 06/15/10 None Available Tamaraw 06/16/10 06/16/10 None Available Forehead Sichuan Takin 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot West Caucasian Tur 06/15/10 06/15/10 06/15/10 None Available Goat 06/16/10 06/16/10 06/16/10 None Available Alpine Ibex 06/17/10 06/17/10 06/17/10 None Available Mainland Serow 06/18/10 06/18/10 None Available Forehead Zebra Duiker 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Blue Wildebeest 06/23/10 06/23/10 None Available Forehead Red-fronted Gazelle 06/23/10 06/23/10 None Available Forehead Nilgiri Tahr 06/18/10 06/18/10 06/18/10 None Available Roan Antelope 07/02/10 07/02/10 07/02/10 Forehead, Back of Thigh Sable Antelope 07/02/10 07/02/10 07/02/10 None Available Waterbuck 06/23/10 06/23/10 None Available Forehead Gerenuk 06/23/10 06/23/10 None Available Forehead Dik Dik 06/23/10 06/23/10 None Available Forehead Himalayan Goral 07/02/10 07/02/10 07/02/10 None Available 75

Grant's Gazelle 06/23/10 06/23/10 None Available Forehead Mountain Goat 06/23/10 06/23/10 06/23/10 None Available Klipspringer 06/23/10 06/23/10 None Available Forehead Beisa Oryx 06/23/10 06/23/10 None Available Forehead Gemsbok 07/02/10 07/02/10 07/02/10 None Available Muskox 07/02/10 07/02/10 07/02/10 None Available Argali 07/02/10 None Available None Available Top of Foot Marco Polo Sheep 07/02/10 07/02/10 07/02/10 None Available Lincolnshire Cross Sheep 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Mountain Big Horn Sheep 06/23/10 06/23/10 06/23/10 None Available Dall's Sheep 06/23/10 06/23/10 06/23/10 None Available Chiru 07/08/10 07/08/10 None Available Forehead Blue Duiker 07/08/10 07/08/10 07/08/10 None Available Chinese Bharal 07/08/10 07/08/10 07/08/10 None Available Steenbok 06/23/10 06/23/10 None Available Forehead Bohor Reedbuck 06/23/10 06/23/10 None Available Forehead Chamois 07/08/10 07/08/10 None Available Forehead Saiga Antelope 07/08/10 07/08/10 07/08/10 None Available Common Duiker 07/11/10 07/11/10 07/11/10 None Available Cape Buffalo 07/18/10 07/18/10 None Available Forehead Giant Eland 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Lowland Nyala 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Top of Foot Greater Kudu 06/23/10 06/23/10 None Available Forehead Dromedary Camel 07/11/10 07/11/10 07/11/10 None Available Llama 07/11/10 07/11/10 07/11/10 None Available Guanaco 07/11/10 07/11/10 07/11/10 None Available 76

Vicugna 07/18/10 07/18/10 07/18/10 None Available Moose 06/23/10 None Available 06/23/10 Forehead Hog Deer 07/18/10 07/18/10 07/18/10 None Available Roe Deer 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Elk 06/23/10 06/23/10 None Available Forehead Red Deer 07/18/10 07/18/10 07/18/10 None Available Sika Deer 07/11/10 07/11/10 07/18/10 None Available Fallow Deer 07/18/10 07/18/10 None Available Forehead Pere David's Deer 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Siberian Musk Deer 07/11/10 07/11/10 07/11/10 None Available Indian Muntjac 06/23/10 06/23/10 06/23/10 Forehead, Top of Foot, back of thigh, vibrissae Reeve's Muntjac 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Mule Deer 06/23/10 06/23/10 06/23/10 None Available White-tailed Deer 06/05/10 06/05/10 06/05/10 None Available Caribou 06/23/10 06/23/10 06/23/10 None Available Giraffe 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Top of Foot Okapi 06/05/10 06/05/10 06/05/10 Back of Thigh, Forehead Pig 06/23/10 06/23/10 06/23/10 None Available Wild Boar 06/23/10 06/23/10 06/23/10 None Available Collared Peccary 06/23/10 06/23/10 06/23/10 Unspecified Lesser Mouse Deer 06/23/10 06/23/10 06/23/10 None Available Carnivora Red Panda 06/23/10 06/23/10 06/23/10 None Available Golden Jackal 06/23/10 06/23/10 06/23/10 None Available Dog 06/23/10 06/23/10 06/23/10 None Available Coyote 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Grey Wolf 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot 77

Dhole 06/23/10 06/23/10 06/23/10 None Available African Wild Dog 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae Raccoon Dog 06/23/10 06/23/10 06/23/10 None Available Grey Fox 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh Red Fox 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Cheetah 06/23/10 06/23/10 06/23/10 None Available Domestic Cat 06/23/10 06/23/10 06/23/10 None Available Ocelot 06/23/10 06/23/10 06/23/10 None Available Lynx 06/23/10 06/23/10 06/23/10 None Available Bobcat 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Clouded Leopard 06/23/10 06/23/10 06/23/10 None Available Lion 06/23/10 06/23/10 06/23/10 None Available Jaguar 06/23/10 06/23/10 06/23/10 None Available Leopard 06/23/10 06/23/10 06/23/10 None Available Sumatran Tiger 07/15/10 07/15/10 07/15/10 None Available Bengal Tiger 07/15/10 07/15/10 07/15/10 None Available Leopard Cat 07/15/10 07/15/10 None Available Forehead Puma 09/13/10 09/13/10 09/13/10 None Available Jaguarundi 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Snow Leopard 09/13/10 09/13/10 09/13/10 None Available African Meerkat None Avalable 07/15/10 07/15/10 Forehead Spotted Hyena 06/23/10 06/23/10 06/23/10 None Available Brown Hyena 07/15/10 07/15/10 07/15/10 None Available Striped Skunk 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Eastern Spotted Skunk 07/15/10 07/15/10 07/15/10 None Available Oriental small clawed otter 06/23/10 06/23/10 06/23/10 None Available 78

Sea Otter 07/15/10 07/15/10 07/15/10 None Available Wolverine 08/01/10 08/01/10 08/01/10 None Available North American Otter 07/15/10 07/15/10 07/15/10 None Available American Marten 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh Fisher 08/01/10 08/01/10 08/01/10 Top of foot, Forehead, Vibrissae, Back of thigh Sable 08/01/10 08/01/10 08/01/10 None Available Ermine 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Long-tailed Weasel 06/23/10 06/23/10 06/23/10 Top of foot, Forehead, Vibrissae, Back of thigh Black-footed Ferret 08/01/10 08/01/10 08/01/10 None Available Ferret 08/01/10 08/01/10 08/01/10 None Available Siberian Mink 08/01/10 08/01/10 08/01/10 None Available American Mink 06/23/10 06/23/10 06/23/10 None Available American Badger 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Northern Fur Seal 06/23/10 06/23/10 None Available Forehead Sea Lion 08/01/10 08/01/10 08/01/10 None Available Harbor Seal 07/15/10 07/15/10 None Available Forehead Ringtail Cat 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot White-nosed Coati 06/23/10 06/23/10 06/23/10 None Available Kinkajou 06/23/10 06/23/10 06/23/10 Forehead, Top of foot Raccoon 06/23/10 06/23/10 06/23/10 None Available Giant Panda 06/23/10 06/23/10 06/23/10 None Available Malayan Sunbear 08/01/10 08/01/10 08/01/10 None Available Sloth Bear 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Spectacled Bear 08/01/10 08/01/10 08/01/10 None Available American Black Bear 06/23/10 06/23/10 06/23/10 Back of Thigh, Forehead, Vibrissae, Top of Foot Brown Bear 06/23/10 06/23/10 06/23/10 None Available 79

Grizzly Bear 06/23/10 06/23/10 06/23/10 None Available Polar Bear 08/01/10 08/01/10 08/01/10 None Available Asiatic Black Bear 06/23/10 06/23/10 06/23/10 None Available Genet Cat 08/01/10 08/01/10 08/01/10 None Available

80

APPENDIX F: HAIRBASE™ IMAGING AND WEBPAGE DESIGN CHECKLIST

Order Common Dorsal Ventral Tip of Tail Other Body Part Images Images Images Webpage SEM Image Name Images Images Taken Images Taken Images Taken Photoshopped Uploaded Checked Complete Taken Taken Artiodactyla Pronghorn 10/11/10 10/11/10 None Available Forehead 10/14/10 10/14/10 10/18/10 10/18/10 Antelope 06/23/10 Addax 10/11/10 10/11/10 10/11/10 None None 10/14/10 10/14/10 10/18/10 10/18/10 Available Impala 10/11/10 10/11/10 None Available Forehead 10/14/10 10/14/10 10/18/10 10/18/10 06/23/10 Kongoni 10/11/10 10/11/10 None Available Forehead None 10/14/10 10/14/10 10/18/10 10/18/10 Hartebeest Available Springbok 10/11/10 10/11/10 None Available Forehead None 10/14/10 10/14/10 10/18/10 10/18/10 Available American 10/12/10 10/12/10 10/12/10 None 10/15/10 10/15/10 10/18/10 10/18/10 Bison 06/23/10 Yak 10/12/10 10/12/10 10/12/10 None None 10/15/10 10/15/10 10/18/10 10/18/10 Available Java 10/12/10 10/12/10 10/12/10 None None 10/15/10 10/15/10 10/18/10 10/18/10 Banteng Available Domestic 10/12/10 10/12/10 10/12/10 None 10/15/10 10/15/10 10/18/10 10/18/10 Cow 06/23/10 Tamaraw 10/12/10 10/12/10 None Available Forehead None 10/15/10 10/15/10 10/18/10 10/18/10 Available Sichuan 10/12/10 10/12/10 10/12/10 None None 10/15/10 10/15/10 10/18/10 10/18/10 Takin Available West 10/12/10 10/12/10 10/12/10 None 10/15/10 10/15/10 10/18/10 10/18/10 Caucasian None Tur Available Goat 10/18/10 10/18/10 10/18/10 None None 10/21/10 10/21/10 10/25/10 10/25/10 Available Alpine Ibex 10/18/10 10/18/10 10/18/10 None None 10/21/10 10/21/10 10/25/10 10/25/10 Available Mainland 10/18/10 10/18/10 None Available Forehead None 10/21/10 10/21/10 10/25/10 10/25/10 Serow Available Zebra 10/18/10 10/18/10 10/18/10 None None 10/21/10 10/21/10 10/25/10 10/25/10 Duiker Available Blue 10/18/10 10/18/10 None Available Forehead None 10/21/10 10/21/10 10/25/10 10/25/10 Wildebeest Available 82

Red-fronted 10/21/10 10/21/10 None Available Forehead 10/22/10 10/22/10 10/25/10 10/25/10 Gazelle 06/23/10 Nilgiri Tahr 10/21/10 10/21/10 10/21/10 None None 10/22/10 10/22/10 10/25/10 10/25/10 Available Roan 10/21/10 10/21/10 10/21/10 None None 10/22/10 10/22/10 10/25/10 10/25/10 Antelope Available Sable 10/21/10 10/21/10 10/21/10 None None 10/22/10 10/22/10 10/25/10 10/25/10 Antelope Available Waterbuck 10/21/10 10/21/10 None Available Forehead None 10/22/10 10/22/10 10/25/10 10/25/10 Available Gerenuk 10/21/10 10/21/10 None Available Forehead None 10/22/10 10/22/10 10/25/10 10/25/10 Available Dik Dik 11/08/10 11/08/10 None Available Forehead 11/11/10 11/11/10 11/15/10 11/15/10 06/23/10 Himalayan 11/08/10 11/08/10 11/08/10 None None 11/11/10 11/11/10 11/15/10 11/15/10 Goral Available Grant's 11/08/10 11/08/10 None Available Forehead 11/11/10 11/11/10 11/15/10 11/15/10 Gazelle 06/23/10 Mountain 11/08/10 11/08/10 11/08/10 None 11/11/10 11/11/10 11/15/10 11/15/10 Goat 06/23/10 Klipspringer 11/08/10 11/08/10 None Available Forehead None 11/11/10 11/11/10 11/15/10 11/15/10 Available Beisa Oryx 11/08/10 11/08/10 None Available Forehead None 11/11/10 11/11/10 11/15/10 11/15/10 Available Gemsbok 11/09/10 11/09/10 11/09/10 None None 11/12/10 11/12/10 11/15/10 11/15/10 Available Muskox 11/09/10 11/09/10 11/09/10 None None 11/12/10 11/12/10 11/15/10 11/15/10 Available Argali 11/09/10 None Available None Available Top of Foot None 11/12/10 11/12/10 11/15/10 11/15/10 Available Marco Polo 11/09/10 11/09/10 11/09/10 None None 11/12/10 11/12/10 11/15/10 11/15/10 Sheep Available Lincolnshire 11/09/10 11/09/10 11/09/10 None None 11/12/10 11/12/10 11/15/10 11/15/10 Cross Sheep Available Mountain 11/18/10 11/18/10 11/18/10 None 11/19/10 11/19/10 11/22/10 11/22/10 Big Horn Sheep 06/23/10 Dall's Sheep 11/18/10 11/18/10 11/18/10 None None 11/19/10 11/19/10 11/22/10 11/22/10 Available Chiru 11/18/10 11/18/10 None Available Forehead None 11/19/10 11/19/10 11/22/10 11/22/10 Available 83

Blue Duiker 11/18/10 11/18/10 11/18/10 None 11/19/10 11/19/10 11/22/10 11/22/10 06/23/10 Chinese 11/18/10 11/18/10 11/18/10 None None 11/19/10 11/19/10 11/22/10 11/22/10 Bharal Available Steenbok 11/18/10 11/18/10 None Available Forehead 11/19/10 11/19/10 11/22/10 11/22/10 06/23/10 Bohor 11/18/10 11/18/10 None Available Forehead 11/19/10 11/19/10 11/22/10 11/22/10 Reedbuck 06/23/10 Chamois 12/06/10 12/06/10 None Available Forehead None 12/09/10 12/09/10 01/10/11 01/10/11 Available Saiga 12/06/10 12/06/10 12/06/10 None None 12/09/10 12/09/10 01/10/11 01/10/11 Antelope Available Common 12/06/10 12/06/10 12/06/10 None None 12/09/10 12/09/10 01/10/11 01/10/11 Duiker Available Cape 12/06/10 12/06/10 None Available Forehead None 12/09/10 12/09/10 01/10/11 01/10/11 Buffalo Available Giant Eland 12/06/10 12/06/10 12/06/10 None None 12/09/10 12/09/10 01/10/11 01/10/11 Available Lowland 12/06/10 12/06/10 12/06/10 None None 12/09/10 12/09/10 01/10/11 01/10/11 Nyala Available Greater 12/07/10 12/07/10 None Available Forehead 12/10/11 12/10/11 01/10/11 01/10/11 Kudu 06/23/10 Dromedary 12/07/10 12/07/10 12/07/10 None None 12/10/11 12/10/11 01/10/11 01/10/11 Camel Available Llama 12/07/10 12/07/10 12/07/10 None None 12/10/11 12/10/11 01/10/11 01/10/11 Available Guanaco 12/07/10 12/07/10 12/07/10 None None 12/10/11 12/10/11 01/10/11 01/10/11 Available Vicugna 12/07/10 12/07/10 12/07/10 None None 12/10/11 12/10/11 01/10/11 01/10/11 Available Moose 12/07/10 None Available 12/07/10 Forehead 12/10/11 12/10/11 01/10/11 01/10/11 06/23/10 Hog Deer 12/07/10 12/07/10 12/07/10 None None 12/10/11 12/10/11 01/10/11 01/10/11 Available Roe Deer 12/07/10 12/07/10 12/07/10 None None 12/10/11 12/10/11 01/10/11 01/10/11 Available Elk 12/13/10 01/03/11 None Available Forehead 01/06/11 01/06/11 01/17/11 01/17/11 06/23/10 Red Deer 12/13/10 01/03/11 01/03/11 None None 01/06/11 01/06/11 01/17/11 01/17/11 Available Sika Deer 12/13/10 01/03/11 01/03/11 None None 01/06/11 01/06/11 01/17/11 01/17/11 Available 84

Fallow Deer 12/13/10 01/03/11 None Available Forehead None 01/06/11 01/06/11 01/17/11 01/17/11 Available Pere David's 12/13/10 01/03/11 01/03/11 None None 01/06/11 01/06/11 01/17/11 01/17/11 Deer Available Siberian 12/13/10 01/03/11 01/03/11 None None 01/06/11 01/06/11 01/17/11 01/17/11 Musk Deer Available Indian 12/13/10 01/03/11 01/03/11 None None 01/06/11 01/06/11 01/17/11 01/17/11 Muntjac Available Reeve's 12/14/10 01/04/11 01/04/10 None None 01/07/11 01/07/11 01/17/11 01/17/11 Muntjac Available Mule Deer 12/14/10 01/04/11 01/04/10 None 01/07/11 01/07/11 01/17/11 01/17/11 06/23/10 White-tailed 12/14/10 01/04/11 01/04/10 None None 01/07/11 01/07/11 01/17/11 01/17/11 Deer Available Caribou 12/14/10 01/04/11 01/04/10 None None 01/07/11 01/07/11 01/17/11 01/17/11 Available Giraffe 12/14/10 01/04/11 01/04/10 None None 01/07/11 01/07/11 01/17/11 01/17/11 Available Okapi 12/14/10 01/04/11 01/04/10 None None 01/07/11 01/07/11 01/17/11 01/17/11 Available Pig 12/14/10 01/04/11 01/04/10 None None 01/07/11 01/07/11 01/17/11 01/17/11 Available Wild Boar 12/14/10 01/04/11 01/04/10 None None 01/07/11 01/07/11 01/17/11 01/17/11 Available Collared 12/15/10 01/10/11 01/10/11 Unspecified None 01/13/11 01/13/11 01/17/11 01/17/11 Peccary Available Lesser 12/15/10 01/10/11 01/10/11 None None 01/13/11 01/13/11 02/07/11 02/07/11 Mouse Deer Available Carnivora Red Panda 12/15/10 01/10/11 01/10/11 None 01/13/11 01/13/11 02/07/11 02/07/11 06/23/10 Golden 12/16/10 01/10/11 01/10/11 None None 01/13/11 01/13/11 02/07/11 02/07/11 Jackal Available Dog 12/16/10 01/10/11 01/10/11 None None 01/13/11 01/13/11 02/07/11 02/07/11 Available Coyote 12/16/10 01/10/11 01/10/11 None 01/13/11 01/13/11 02/07/11 02/07/11 06/23/10 Grey Wolf 12/16/10 01/10/11 01/10/11 None 01/13/11 01/13/11 02/07/11 02/07/11 06/23/10 Dhole 09/07/10 09/07/10 09/07/10 None None 09/09/10 09/09/10 09/13/10 09/13/10 Available African 09/07/10 09/07/10 09/07/10 None None 09/09/10 09/09/10 09/13/10 09/13/10 Wild Dog Available 85

Raccoon 09/07/10 09/07/10 09/07/10 None None 09/09/10 09/09/10 09/13/10 09/13/10 Dog Available Grey Fox 09/07/10 09/07/10 09/07/10 None None 09/09/10 09/09/10 09/13/10 09/13/10 Available Red Fox 08/16/10 08/16/10 08/16/10 None 09/09/10 09/09/10 09/13/10 09/13/10 06/23/10 Cheetah 09/07/10 09/07/10 09/07/10 None 09/09/10 09/09/10 09/13/10 09/13/10 06/23/10 Domestic 09/07/10 09/07/10 09/07/10 None None 09/09/10 09/09/10 09/13/10 09/13/10 Cat Available Ocelot 08/16/10 08/16/10 08/16/10 None None 09/09/10 09/09/10 09/13/10 09/13/10 Available Lynx 09/08/10 09/08/10 09/08/10 None None 09/09/10 09/09/10 09/13/10 09/13/10 Available Bobcat 09/08/10 09/08/10 09/08/10 None 09/09/10 09/09/10 09/13/10 09/13/10 06/23/10 Clouded 09/08/10 09/08/10 09/08/10 None None 09/09/10 09/09/10 09/13/10 09/13/10 Leopard Available Lion 08/16/10 08/16/10 08/16/10 None 09/09/10 09/09/10 09/13/10 09/13/10 06/23/10 Jaguar 09/13/10 09/13/10 09/13/10 None None 09/16/10 09/16/10 09/20/10 09/20/10 Available Leopard 09/13/10 09/13/10 09/13/10 None 09/16/10 09/16/10 09/20/10 09/20/10 06/23/10 Sumatran 09/13/10 09/13/10 09/13/10 None None 09/16/10 09/16/10 09/20/10 09/20/10 Tiger Available Bengal 09/13/10 09/13/10 09/13/10 None None 09/16/10 09/16/10 09/20/10 09/20/10 Tiger Available Leopard Cat 08/16/10 08/16/10 None Available Forehead None 09/16/10 09/16/10 09/20/10 09/20/10 Available Puma 09/14/10 09/14/10 09/14/10 None None 09/17/10 09/17/10 09/20/10 09/20/10 Available Jaguarundi 09/14/10 09/14/10 09/14/10 None None 09/17/10 09/17/10 09/20/10 09/20/10 Available Snow 09/14/10 09/14/10 09/14/10 None None 09/17/10 09/17/10 09/20/10 09/20/10 Leopard Available African None 09/14/10 09/14/10 Forehead None 09/17/10 09/17/10 09/20/10 09/20/10 Meerkat Available Available Spotted 09/14/10 09/14/10 09/14/10 None None 09/17/10 09/17/10 09/20/10 09/20/10 Hyena Available Brown 09/14/10 09/14/10 09/14/10 None None 09/17/10 09/17/10 09/20/10 09/20/10 Hyena Available 86

Striped 08/16/10 08/16/10 08/16/10 None 09/17/10 09/17/10 09/20/10 09/20/10 Skunk 06/23/10 Eastern 09/20/10 09/20/10 09/20/10 None 09/23/10 09/23/10 09/27/10 09/27/10 Spotted None Skunk Available Oriental 09/20/10 09/20/10 09/20/10 None 09/23/10 09/23/10 09/27/10 09/27/10 small None clawed otter Available Sea Otter 09/20/10 09/20/10 09/20/10 None None 09/23/10 09/23/10 09/27/10 09/27/10 Available Wolverine 09/20/10 09/20/10 09/20/10 None None 09/23/10 09/23/10 09/27/10 09/27/10 Available North 09/20/10 09/20/10 09/20/10 None 09/23/10 09/23/10 09/27/10 09/27/10 American None Otter Available American 09/20/10 09/20/10 09/20/10 None None 09/23/10 09/23/10 09/27/10 09/27/10 Marten Available Fisher 09/20/10 09/20/10 09/20/10 None None 09/23/10 09/23/10 09/27/10 09/27/10 Available Sable 08/16/10 08/16/10 08/16/10 None None 09/24/10 09/24/10 09/27/10 09/27/10 Available Ermine 09/21/10 09/21/10 09/21/10 None 09/24/10 09/24/10 09/27/10 09/27/10 06/23/10 Long-tailed 09/21/10 09/21/10 09/21/10 None 09/24/10 09/24/10 09/27/10 09/27/10 Weasel 06/23/10 Black-footed 09/21/10 09/21/10 09/21/10 None None 09/24/10 09/24/10 09/27/10 09/27/10 Ferret Available Ferret 09/21/10 09/21/10 09/21/10 None None 09/24/10 09/24/10 09/27/10 09/27/10 Available Siberian 09/21/10 09/21/10 09/21/10 None None 09/24/10 09/24/10 09/27/10 09/27/10 Mink Available American 09/21/10 09/21/10 09/21/10 None 09/24/10 09/24/10 09/27/10 09/27/10 Mink 06/23/10 American 09/21/10 09/21/10 09/21/10 None 09/24/10 09/24/10 09/27/10 09/27/10 Badger 06/23/10 Northern 08/16/10 08/16/10 None Available Forehead None 09/24/10 09/24/10 09/27/10 09/27/10 Fur Seal Available Sea Lion 09/27/10 09/27/10 09/27/10 None 09/30/10 09/30/10 10/04/10 10/04/10 06/23/10 Harbor Seal 09/27/10 09/27/10 None Available Forehead 09/30/10 09/30/10 10/04/10 10/04/10 06/23/10 87

Ringtail Cat 09/27/10 09/27/10 09/27/10 None None 09/30/10 09/30/10 10/04/10 10/04/10 Available White-nosed 09/27/10 09/27/10 09/27/10 None 09/30/10 09/30/10 10/04/10 10/04/10 Coati 06/23/10 Kinkajou 09/27/10 09/27/10 09/27/10 None None 09/30/10 09/30/10 10/04/10 10/04/10 Available Raccoon 09/27/10 09/27/10 09/27/10 None 09/30/10 09/30/10 10/04/10 10/04/10 06/23/10 Giant Panda 09/27/10 09/27/10 09/27/10 None None 09/30/10 09/30/10 10/04/10 10/04/10 Available Malayan 08/16/10 08/16/10 08/16/10 None None 09/30/10 09/30/10 10/04/10 10/04/10 Sunbear Available Sloth Bear 09/28/10 09/28/10 09/28/10 None None 10/01/10 10/01/10 10/04/10 10/04/10 Available Spectacled 09/28/10 09/28/10 09/28/10 None None 10/01/10 10/01/10 10/04/10 10/04/10 Bear Available American 09/28/10 09/28/10 09/28/10 None None 10/01/10 10/01/10 10/04/10 10/04/10 Black Bear Available Brown Bear 09/28/10 09/28/10 09/28/10 None 10/01/10 10/01/10 10/04/10 10/04/10 06/23/10 Grizzly Bear 09/28/10 09/28/10 09/28/10 None None 10/01/10 10/01/10 10/04/10 10/04/10 Available Polar Bear 09/28/10 09/28/10 09/28/10 None 10/01/10 10/04/10 10/04/10 10/4/10 06/23/10 Asiatic 08/16/10 08/16/10 08/16/10 None 10/01/10 10/04/10 10/04/10 10/4/10 Black Bear 06/23/10 Genet Cat 09/28/10 09/28/10 09/28/10 None 10/01/10 10/04/10 10/04/10 10/4/10 06/23/10

88 California State University, Fresno

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3/25/11

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